Secreted and transmembrane polypeptides and nucleic acids encoding the same

ABSTRACT

The present invention is directed to novel polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

FIELD OF THE INVENTION

[0001] The present invention relates generally to the identification andisolation of novel DNA and to the recombinant production of novelpolypeptides.

BACKGROUND OF THE INVENTION

[0002] Extracellular proteins play important roles in, among otherthings, the formation, differentiation and maintenance of multicellularorganisms. The fate of many individual cells, e.g., proliferation,migration, differentiation, or interaction with other cells, istypically governed by information received from other cells and/or theimmediate environment. This information is often transmitted by secretedpolypeptides (for instance, mitogenic factors, survival factors,cytotoxic factors, differentiation factors, neuropeptides, and hormones)which are, in turn, received and interpreted by diverse cell receptorsor membrane-bound proteins. These secreted polypeptides or signalingmolecules normally pass through the cellular secretory pathway to reachtheir site of action in the extracellular environment.

[0003] Secreted proteins have various industrial applications, includingas pharmaceuticals, diagnostics, biosensors and bioreactors. Mostprotein drugs available at present, such as thrombolytic agents,interferons, interleukins, erythropoietins, colony stimulating factors,and various other cytokines, are secretory proteins. Their receptors,which are membrane proteins, also have potential as therapeutic ordiagnostic agents. Efforts are being undertaken by both industry andacademia to identify new, native secreted proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. Examples ofscreening methods and techniques are described in the literature [see,for example, Klein et al., Proc. Natl. Acad. Sci. 93:7108-7113 (1996);U.S. Pat. No. 5,536,637)].

[0004] Membrane-bound proteins and receptors can play important rolesin, among other things, the formation, differentiation and maintenanceof multicellular organisms. The fate of many individual cells, e.g.,proliferation, migration, differentiation, or interaction with othercells, is typically governed by information received from other cellsand/or the immediate environment. This information is often transmittedby secreted polypeptides (for instance, mitogenic factors, survivalfactors, cytotoxic factors, differentiation factors, neuropeptides, andhormones) which are, in turn, received and interpreted by diverse cellreceptors or membrane-bound proteins. Such membrane-bound proteins andcell receptors include, but are not limited to, cytokine receptors,receptor kinases, receptor phosphatases, receptors involved in cell-cellinteractions, and cellular adhesin molecules like selectins andintegrins. For instance, transduction of signals that regulate cellgrowth and differentiation is regulated in part by phosphorylation ofvarious cellular proteins. Protein tyrosine kinases, enzymes thatcatalyze that process, can also act as growth factor receptors. Examplesinclude fibroblast growth factor receptor and nerve growth factorreceptor.

[0005] Membrane-bound proteins and receptor molecules have variousindustrial applications, including as pharmaceutical and diagnosticagents. Receptor immunoadhesins, for instance, can be employed astherapeutic agents to block receptor-ligand interactions. Themembrane-bound proteins can also be employed for screening of potentialpeptide or small molecule inhibitors of the relevant receptor/ligandinteraction.

[0006] Efforts are being undertaken by both industry and academia toidentify new, native receptor or membrane-bound proteins. Many effortsare focused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel receptor or membrane-boundproteins.

[0007] 1. PRO1484

[0008] Adipose differentiation is accompanied by changes in cellularmorphology, a dramatic accumulation of intracellular lipid andactivation of a specific program of gene expression (Liang et al., J.Biol. Chem. 271:10697-10703 (1996)). Adipose complement-related proteinis a protein whose expression is highly induced during adipocytedifferentiation and which shares significant homology with subunits ofcomplement factor C1q, collagen alpha 1(x) and the brain-specific factocerebellin (Scherer et al., J. Biol. Chem. 270:26746-26749 (1995)).While the function of adipocyte complement-related protein is presentlyunknown, the tissue-specific expression thereof suggests that thisprotein functions as a novel signaling molecule for adipose tissue. Assuch, there is significant interest in identifying and characterizingnovel polypeptides having homology to adipocyte complement-relatedprotein. We herein describe the identification and characterization ofnovel polypeptides having homology to adipocyte complement-relatedprotein, designated herein as PRO1484 polypeptides.

[0009] 2. PRO4334

[0010] Plasma cell membrane glycoprotein PC-1 is of interest. Thecloning of PC-1 is described in the art, i.e., see Buckley, et al., J.Biol. Chem., 265(29):17506-11 (1990) and WO9519570-A. WO9519570-Adescribes the human insulin receptor tyrosine kinase inhibitor PC-1. Itis reported that PC-1 is useful in the diagnosis and treatment ofdiseases involving inappropriate insulin receptor tyrosine kinaseinhibitor expression such as non-insulin dependent mellitus. Thus,proteins having homology to PC-1 are of interest.

[0011] 3. PRO1122

[0012] It has been reported that the cytokine interleukin 17 (IL-17)stimulates epithelial, endothelial, and fibroblastic cells to secretecytokines such as IL-6, IL-8, and granulocyte-colony-stimulating factor,as well as prostaglandin E2. Moreover, it has been shown that whencultured in the presence of IL-17, fibroblasts could sustainproliferation of CD34+ preferential maturation into neutrophils. Thus ithas been suggested that IL-17 constitutes an early initiator of the Tcelldependent inflammatory reaction and/or an element of the cytokinenetwork that bridges the immune system to hematopoiesis. See, Yao, etal., J. Immunol., 155(12):5483-5486 (1995); Fossiez, et al., J. Exp.Med., 183(6):2593-2603 (1996); Kennedy, et al., J. Interferon CytokineRes., 16(8):611-617 (1996). Thus, proteins related to IL-17, includingCTLA-8, which has been mistaken for IL-17 (see Kennedy, supra) are ofinterest.

[0013] 4. PRO1889

[0014] E48 antigen protein is a cysteine-rich GPI-anchored membraneprotein that belongs to the LY-6 family of human proteins (see, e.g., WO96/35808). The E48 antigen serves as a marker for squamous cells,exhibits biological activity of cell-cell/cell-matrix adhesion and is atarget for antibody-based immunotherapy. The amino acid sequence of theE48 antigen protein has previously been deduced from a cDNA clone whichwas obtained from a squamous cell carcinoma of the head and neck. Assuch, the E48 antigen serves as a potential target for the treatment ofsquamous cell cancer.

[0015] We herein describe the identification and characterization ofnovel polypeptides having homology to E48 antigen protein, designatedherein as PRO1889 polypeptides.

[0016] 5. PRO1890

[0017] The recognition of carbohydrates by lectins has been found toplay an important role in various aspects of eukaryotic physiology. Anumber of different animal and plant lectin families exist, but it isthe calcium dependent, or type C, lectins that have recently garneredthe most attention. For example, the recognition of carbohydrateresidues on either endothelial cells or leukocytes by the selectinfamily of calcium dependent lectins has been found to be of profoundimportance to the trafficking of leukocytes to inflammatory sites.Lasky, L., Ann. Rev. Biochem., 64 113-139 (1995). The biophysicalanalysis of these adhesive interactions has suggested thatlectin-carbohydrate binding evolved in this case to allow for theadhesion between leukocytes and the endothelium under the high shearconditions of the vasculature. Thus, the rapid on rates of carbohydraterecognition by such lectins allows for a hasty acquisition of ligand, anecessity under the high shear of the vascular flow. The physiologicaluse of type C lectins in this case is also supported by the relativelylow affinities of these interactions, a requirement for the leukocyterolling phenomenon that has been observed to occur at sites of acuteinflammation. The crystal structures of the mannose binding protein(Weis et al., Science 254, 1608-1615 [1991]; Weis et al., Nature 360127-134 [1992]) and E-selectin (Graves et al., Nature 367(6463), 532-538[1994]), together with various mutagenesis analyses (Erbe et al., J.Cell. Biol. 119(1), 215-227 [1992]; Drickamer, Nature 360, 183-186[1992]; Iobst et al., J. Biol. Chem. 169(22), 15505-15511 [1994]; Koganet al., J. Biol. Chem. 270(23), 14047-14055 [1995]), is consistent withthe supposition that the type C lectins are, in general, involved withthe rapid recognition of clustered carbohydrates. Together, these datasuggest that type C lectins perform a number of critical physiologicalphenomena through the rapid, relatively low affinity recognition ofcarbohydrates.

[0018] Given the obvious importance of the lectin proteins in numerousbiological processes, efforts are currently being made to identify novellectin proteins or proteins having sequence homology to lectin proteins.We herein describe the identification and characterization of novelpolypeptides having homology to a lectin protein, designated herein asPRO1890 polypeptides.

[0019] 6. PRO1887

[0020] Enzymatic proteins play important roles in the chemical reactionsinvolved in the digestion of foods, the biosynthesis of macromolecules,the controlled release and utilization of chemical energy, and otherprocesses necessary to sustain life. Enzymes have also been shown toplay important roles in combating various diseases and disorders. Forexample, liver carboxylesterases have been reported to assist insensitizing human tumor cells to the cancer prodrugs. Danks et al.,report that stable expression of the cDNA encoding a carboxylesterase inRh30 human rhabdomyosarcoma cells increased the sensitivity of the cellsto the CPT-11 cancer prodrug 8.1-fold. Cancer Res. (1998) 58(1):20-22.The authors propose that this prodrug/enzyme combination could beexploited therapeutically in a manner analogous to approaches currentlyunder investigation with the combinations of ganciclovir/herpes simplexvirus thymidine kinase and 5-fluorocytosine/cytosine deaminase. van Peltet al. demonstrated that a 55 kD human liver carboxylesterase inhibitsthe invasion of Plasmodium falciparum malaria sporozoites into primaryhuman hepatocytes in culture. J Hepatol (1997) 27(4):688-698.

[0021] Carboxylesterases have also been found to be of importance in thedetoxification of drugs, pesticides and other xenobiotics. Purifiedhuman liver carboxylesterases have been shown to be involved in themetabolism of various drugs including cocaine and heroin. Prindel et al.describe the purification and cloning of a broad substrate specificityhuman liver carboxylesterase which catalyzes the hydrolysis of cocaineand heroin and which may play an important role in the degradation ofthese drugs in human tissues. J. Biol. Chem. (1997)6:272(23):14769-14775. Brzenzinski et al. describe a spectrophotometriccompetitive inhibition assay used to identify drug or environmentalesters that are metabolized by carboxylesterases. Drug Metab Dispos(1997) 25(9):1089-1096.

[0022] As additional background information on carboxylesterases, Kroetzet al. (Biochemistry, (1993) 32(43): 11606-17) reported the cDNA cloningand characterization of human liver carboxylesterases. Aida et al.(Biochim Biophys Acta (1993) 1174(l):72-4) reported the cDNA cloning andcharacterization of a male-predominant carboxylesterase in mouse livercarboxylesterases.

[0023] In light of the important physiological roles played bycarboxylesterases, efforts are being undertaken by both industry andacademia to identify new, native carboxylesterase homologs. We hereindescribe the identification and characterization of a novel polypeptidehaving homology to carboxylesterase.

[0024] 7. PRO1785

[0025] Antioxidant enzymes are thought to play a crucial role in thesurvival of the parasite, Schistosoma mansoni, during its migrationthrough the tissues of a definitive host. Recently, one such enzyme,glutathione peroxidase was cloned. Roche, et al., Gene, 138:149-152(1994), accession number GSHC SCHMA. Glutathione perxodiases are furtherdescribed in FR2689906-A. Thus, glutathione peroxidases, and the nucleicacids which encode them are useful as diagnostic reagents, vaccines andin assays to find modulators of antioxidant enzymes.

[0026] 8. PRO4353

[0027] Semaphorins comprise a large family of proteins implicated inaxonal guidance during development. Semaphorin Y may be used to inhibitperipheral nerve growth. Semaphorin Z is useful as a central nerveextension inhibitor. Semaphorin Z inhibitors can be used as promoters ofcentral nerve regeneration. Thus semaphorins and regulators ofsemaphorins are of great interest. Kikuchi, et al., Brain Res Mol BrainRes., 51(1-2):229-37 (1997); Shoji, et al., Development, 125(7):1275-83(1998).

[0028] 9. PRO4357

[0029] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4357 polypeptides.

[0030] 10. PRO4405

[0031] Efforts are being undertaken by both industry and academia toidentify new, native transmembrane receptor proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel transmembrane receptor proteins.We herein describe the identification and characterization of noveltransmembrane polypeptides, designated herein as PRO4405 polypeptides.

[0032] 11. PRO4356

[0033] Glycosylphosphatidylinositol (GPI) anchored proteoglycans aregenerally localized to the cell surface and are thus known to beinvolved in the regulation of responses of cells to numerous growthfactors, cell adhesion molecules and extracellular matrix components.The metastasis-associated GPI-anchored protein (MAGPIAP) is one of thesecell surface proteins which appears to be involved in metastasis.Metastasis is the form of cancer wherein the transformed or malignantcells are traveling and spreading the cancer from one site to another.Therefore, identifying the polypeptides related to metastasis andMAGPIAP is of interest.

[0034] 12. PRO4352

[0035] Cadherins are a large family of transmembrane proteins. Cadherinscomprise a family of calcium-dependent glycoproteins that function inmediating cell-cell adhesion in virtually all solid tissues ofmulticellular organisms. At least cadherins 1-13 as well as types B, E,EP, M, N, P and R have been characterized. Among the functions cadherinsare known for, with some exceptions, cadherins participate in cellaggregation and are associated with cell-cell adhesion sites. Recently,it has been reported that while all cadherins share multiple repeats ofa cadherin specific motif believed to correspond to folding ofextracellular domains, members of the cadherin superfamily havedivergent structures and, possibly, functions. In particular it has beenreported that members of the cadherin superfamily are involved in signaltransduction. See, Suzuki, J. Cell Biochem., 61(4):531-542 (1996).Cadherins are further described in Tanihara, et al., J. Cell Sci.,107(6):1697-1704 (1994), Aberle, et al., J. Cell Biochem., 61(4):514-523(1996), Obata, et al., Cell Adhes. Commun., 6(4):323-33 (1998) andTanihara, et al., Cell Adhes. Commun., 2(l):15-26 (1994).

[0036] Protocadherins are members of the cadherin superfamily which arehighly expressed in the brain. In some studies, protocadherins haveshown cell adhesion activity. See, Sano, et al., EMBO J.,12(6):2249-2256 (1993). However, studies have also shown that someprotocadherins, such as protocadherin 3 (also referred to as Pcdh3 orpc3), do not show strong calcium dependent cell aggregation activity.See, Sago, et al., Genomics, 29(3):631-640 (1995) for this study andfurther characteristics of Pcdh3. Molecules related to pc3 are thus ofgreat interest. Also of great interest is the subtype of desmosomalcadherin described in Koch, et al., Differentiation, 47(1):29-36 (1991).

[0037] Of particular interest are proteins having a sequence withhomology to that described in Amagai, et al., Cell, 67(5):869-77 (1991).This study describes antibodies against a novel epithelial cadherin inpemphigus vulgaris, a disease of cell adhesion. Also of interest arefull-length cadherins. Additionally, proteins having homology to the fattumor suppressor gene which are novel cadherins are of interest.

[0038] 13. PRO4380

[0039] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4380 polypeptides.

[0040] 14. PRO4354

[0041] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4354 polypeptides.

[0042] 15. PRO4408

[0043] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4408 polypeptides.

[0044] 16. PRO5737

[0045] Interleukin-1 refers to two proteins (IL-1α and IL-1β) which playa key role early in the inflammatory response (for a review, seeDinarello, Blood, 87: 2095-2147 (1996) and references therein). Bothproteins are made as intracellular precursor proteins which are cleavedupon secretion to yield mature carboxy-terminal 17 kDa fragments whichare biologically active. In the case of IL-1β, this cleavage involves anintracellular cysteine protease, known as ICE, which is required torelease the active fragment from the inactive precursor. The precursorof IL-1α is active.

[0046] These two proteins act by binding to cell surface receptors foundon almost all cell types and triggering a range of responses eitheralone or in concert with other secreted factors. These range fromeffects on proliferation (e.g. fibroblasts, T cells) apoptosis (e.g.A375 melanoma cells), cytokine induction (e.g. of TNF, IL-I, IL-8),receptor activation (e.g. E-selectin), eicosanoid production (e.g. PGE2)and the secretion of degradative enzymes (e.g. collagenase). To achievethese effects, IL-1 activates transcription factors such as NF-KB andAP-1. Several of the activities of IL-1 action on target cells arebelieved to be mediated through activation of kinase cascades that havealso been associated with cellular stresses, such as the stressactivated MAP kinase JNK/SAPK and p38.

[0047] A third member of the IL-1 family was subsequently discoveredwhich acts as a natural antagonist of IL-1α and IL-1β by binding to theIL-1 receptor but not transducing an intracellular signal or abiological response. The protein is called IL-1Ra (for IL-1 receptorantagonist) or IRAP (for IL-1 receptor antagonist protein). At leastthree alternatively spliced forms of IL-1Ra exist: one encodes secretedprotein, and the other two encode intracellular proteins. IL-1α, IL-1βand IL-1Ra exhibit approximately 25-30% sequence identity with eachother and share a similar three dimensional structure consisting oftwelve β-strands folded into a β-barrel, with an internal thricerepeated structural motif.

[0048] There are three known IL-1 receptor subunits. The active receptorcomplex consists of the type I receptor and IL-1 accessory protein(IL-1RAcP). The type I receptor is responsible for binding of the IL-1α,IL-1β and IL-1Ra ligands, and is able to do so in the absence of theIL-1RAcP. However, signal transduction requires the interaction of IL-1αor IL-1β with the IL-1RAcP. IL-1Ra does not interact with the IL-1RAcPand hence cannot induce signal transduction. A third receptor subunit,the type II receptor, binds IL-1α and IL-1β but cannot transduce signaldue its lack of an intracellular domain. Instead, the type II receptoreither acts as a decoy in its membrane bound form or as an IL-1antagonist in a processed, secreted form, and hence inhibits IL-1activity. The type II receptor weakly binds to IL-1Ra.

[0049] Many studies using IL-1Ra, soluble IL-1R derived from theextracellular domain of the type I IL-1 receptor, antibodies to IL-1α orIL-1β, and transgenic knockout mice for these genes have shown that IL-1plays a role in a number of pathophysiologies (for a review, seeDinarello, Blood, 87: 2095-2147 (1996)). For example, IL-1Ra has beenshown to be effective in animal models of septic shock, rheumatoidarthritis, graft-versus-host disease (GVHD), stroke, cardiac ischemia,psoriasis, inflammatory bowel disease, and asthma. In addition, IL-1Rahas demonstrated efficacy in clinical trials for rheumatoid arthritisand GVHD, and is also in clinical trials for inflammatory bowel disease,asthma and psoriasis.

[0050] More recently, interleukin-18 (IL-18) was placed in the IL-Ifamily (for a review, see Dinarello et al, J. Leukocyte Biol., 63:658-664 (1998)). IL-18 shares the β-pleated, barrel-like form of IL-1αand IL-1β. In addition, IL-18 is the natural ligand for the IL-1receptor family member formerly known as IL-1R-related protein (IL-1Rrp)(now known as the IL-18 receptor (IL-18R)). IL-18 has been shown toinitiate the inflammatory cytokine cascade in a mixed population ofperipheral blood mononuclear cells (PBMCs) by triggering theconstitutive IL-18 receptors on lymphocytes and NK cells, inducing TNFproduction in the activated cells. TNF, in turn, stimulates IL-1 andIL-8 production in CD14+ cells. Because of its ability to induce TNF,IL-1, and both C—C and C—X—C chemokines, and because IL-18 induces Fasligand as well as nuclear translocation of nuclear factor 6B (NF-6B),IL-18 ranks with other pro-inflammatory cytokines as a likelycontributor to systemic and local inflammation.

[0051] 17. PRO4425

[0052] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4425 polypeptides.

[0053] 18. PRO5990

[0054] The secretogranin proteins (e.g. secretogranin I and II) arefound in secretory granules in a variety of endocrine cells and neurons.Schimmel, A. et al., FEBS Lett. 314(3):375-80 (1992); Gerdes, H. H. etal., J. Biol. Chem. 264(20): 12009-15. A possible function of thesecretogranin proteins is the packaging of secretory products, includingregulatory peptides. Chanat, E. et al., FEBS Lett. 351(2):225-30 (1994);Rosa, P. et al., J. Cell. Biol. 101(5):1999-2011 (1985); Gorr, S. U. etal., Am. J. Physiol. 257(2):E247-54 (1989). Secretogranins have beensuccessfully used as biological markers in a number of contexts. Forexample, secretogranin II has gained importance as animmunohistochemical marker for endocrine neoplasms. See Fischer-Colbrie,R. et al., J. Biol. Chem. 265(16):9208-13 (1990). Eder et al. found thatthe ratio of secretogranin II to chroinogranins was remarkably constantwithin patient populations, and suggest that the ratio may be used as aparameter to standardize CSF levels of other peptides, such asneuropeptides. Eder, U., et al., J. Neural Transm., 105(1):39-51 (1998).

[0055] We herein describe the identification and characterization ofnovel polypeptides having sequence similarity to secretogranin,designated herein as PRO5990 polypeptides.

[0056] 19. PRO6030

[0057] Efforts are being undertaken by both industry and academia toidentify new, native transmembrane receptor proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel transmembrane receptor proteins.We herein describe the identification and characterization of noveltransmembrane polypeptides, designated herein as PRO6030 polypeptides.

[0058] 20. PRO4424

[0059] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4424 polypeptides.

[0060] 21. PRO4422

[0061] Lysozyme is a protein which is widely distributed in severalhuman tissues and secretions including milk, tears and saliva. It hasbeen demonstrated to hydrolyze linkages between N-acetylglucosamines. Ithas been demonstrated to be an inhibitor of chemotaxis and of theproduction of toxic oxygen free radicals and may also have some role inthe calcification process. As such, there is substantial interest inidentifying novel polypeptides having homology to lysozyme. Nakano andGraf, Biochim. Biophys Acta, 1090(2):273-6 (1991).

[0062] 22. PRO4430

[0063] Efforts are being undertaken by both industry and academia toidentify new, native secreted proteins. Many of these efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel secreted proteins. We hereindescribe the identification and characterization of novel secretedpolypeptides, designated herein as PRO4430 polypeptides.

[0064] 23. PRO4499

[0065] Efforts are being undertaken by both industry and academia toidentify new, native transmembrane receptor proteins. Many efforts arefocused on the screening of mammalian recombinant DNA libraries toidentify the coding sequences for novel transmembrane receptor proteins.We herein describe the identification and characterization of noveltransmembrane polypeptides, designated herein as PRO4499 polypeptides.

SUMMARY OF THE INVENTION

[0066] 1. PRO1484

[0067] A cDNA clone (DNA44686-1653) has been identified, having homologyto nucleic acid encoding adipocyte complement-related protein thatencodes a novel polypeptide, designated in the present application as“PRO1484”.

[0068] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1484 polypeptide.

[0069] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO1484 polypeptide having the sequence of aminoacid residues from about 1 or about 23 to about 246, inclusive of FIG. 2(SEQ ID NO:2), or (b) the complement of the DNA molecule of (a).

[0070] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1484 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about nucleotides 77 orabout 143 and about 814, inclusive, of FIG. 1 (SEQ ID NO:1). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0071] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203581 (DNA44686-1653) or (b) the complement of the nucleicacid molecule of (a). In a preferred embodiment, the nucleic acidcomprises a DNA encoding the same mature polypeptide encoded by thehuman protein cDNA in ATCC Deposit No. 203581 (DNA44686-1653).

[0072] In still a further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues 1 or about 23 to about 246, inclusive of FIG. 2 (SEQID NO:2), or (b) the complement of the DNA of (a).

[0073] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least 600 nucleotides and produced byhybridizing a test DNA molecule under stringent conditions with (a) aDNA molecule encoding a PRO1484 polypeptide having the sequence of aminoacid residues from 1 or about 23 to about 246, inclusive of FIG. 2 (SEQID NO:2), or (b) the complement of the DNA molecule of (a), and, if theDNA molecule has at least about an 80% sequence identity, prefereably atleast about an 85% sequence identity, more preferably at least about a90% sequence identity, most preferably at least about a 95% sequenceidentity to (a) or (b), isolating the test DNA molecule.

[0074] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1484 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,or is complementary to such encoding nucleic acid molecule. The signalpeptide has been tentatively identified as extending from about aminoacid position 1 to about amino acid position 22 in the sequence of FIG.2 (SEQ ID NO:2).

[0075] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 1or about 23 to about 246, inclusive of FIG. 2 (SEQ ID NO:2), or (b) thecomplement of the DNA of (a).

[0076] Another embodiment is directed to fragments of a PRO1484polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 to about 80 nucleotides inlength, preferably from about 20 to about 60 nucleotides in length, morepreferably from about 20 to about 50 nucleotides in length and mostpreferably from about 20 to about 40 nucleotides in length and may bederived from the nucleotide sequence shown in FIG. 1 (SEQ ID NO:1).

[0077] In another embodiment, the invention provides isolated PRO1484polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0078] In a specific aspect, the invention provides isolated nativesequence PRO1484 polypeptide, which in certain embodiments, includes anamino acid sequence comprising residues 1 or about 23 to about 246 ofFIG. 2 (SEQ ID NO:2).

[0079] In another aspect, the invention concerns an isolated PRO1484polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues1 or about 23 to about 246, inclusive of FIG. 2 (SEQ ID NO:2).

[0080] In a further aspect, the invention concerns an isolated PRO1484polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 1 orabout 23 to about 246, inclusive of FIG. 2 (SEQ ID NO:2).

[0081] In yet another aspect, the invention concerns an isolated PRO1484polypeptide, comprising the sequence of amino acid residues 1 or about23 to about 246, inclusive of FIG. 2 (SEQ ID NO:2), or a fragmentthereof sufficient to provide a binding site for an anti-PRO1484antibody. Preferably, the PRO1484 fragment retains a qualitativebiological activity of a native PRO1484 polypeptide.

[0082] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1484 polypeptide havingthe sequence of amino acid residues from about 1 or about 23 to about246, inclusive of FIG. 2 (SEQ ID NO: 3), or (b) the complement of theDNA molecule of (a), and if the test DNA molecule has at least about an80% sequence identity, preferably at least about an 85% sequenceidentity, more preferably at least about a 90% sequence identity, mostpreferably at least about a 95% sequence identity to (a) or (b), (ii)culturing a host cell comprising the test DNA molecule under conditionssuitable for expression of the polypeptide, and (iii) recovering thepolypeptide from the cell culture.

[0083] 2. PRO4334

[0084] A cDNA clone (DNA59608-2577) has been identified that encodes anovel polypeptide having homology to PC-1 and designated in the presentapplication as “PRO4334”.

[0085] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4334 polypeptide.

[0086] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4334 polypeptide having the sequence of aminoacid residues from 1 or about 23 to about 440, inclusive of FIG. 4 (SEQID NO:9), or (b) the complement of the DNA molecule of (a).

[0087] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4334 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 150 andabout 1403, inclusive, of FIG. 3 (SEQ ID NO:8). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0088] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203870 (DNA59608-2577), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203870 (DNA59608-2577).

[0089] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 23 to about 440, inclusive of FIG. 4 (SEQID NO:9), or the complement of the DNA of (a).

[0090] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4334 polypeptide having the sequence of amino acid residues fromabout 23 to about 440, inclusive of FIG. 4 (SEQ ID NO:9), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0091] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 23to about 440, inclusive of FIG. 4 (SEQ ID NO:9), or (b) the complementof the DNA of (a).

[0092] Another embodiment is directed to fragments of a PRO4334polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0093] In another embodiment, the invention provides isolated PRO4334polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0094] In a specific aspect, the invention provides isolated nativesequence PRO4334 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 23 through 440 of FIG. 4 (SEQ IDNO:9).

[0095] In another aspect, the invention concerns an isolated PRO4334polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues23 to about 440, inclusive of FIG. 4 (SEQ ID NO:9).

[0096] In a further aspect, the invention concerns an isolated PRO4334polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 23through 440 of FIG. 4 (SEQ ID NO:9).

[0097] In yet another aspect, the invention concerns an isolated PRO4334polypeptide, comprising the sequence of amino acid residues 23 to about440, inclusive of FIG. 4 (SEQ ID NO:9), or a fragment thereof sufficientto provide a binding site for an anti-PRO4334 antibody. Preferably, thePRO4334 fragment retains a qualitative biological activity of a nativePRO4334 polypeptide.

[0098] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4334 polypeptide havingthe sequence of amino acid residues from about 23 to about 440,inclusive of FIG. 4 (SEQ ID NO:9), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0099] 3. PRO1122

[0100] A cDNA clone (DNA62377-1381) has been identified, having sequenceidentity with CTLA-8 that encodes a novel polypeptide, designated in thepresent application as “PRO1122.”In one embodiment, the inventionprovides an isolated nucleic acid molecule comprising DNA encoding aPRO1122 polypeptide.

[0101] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO1122 polypeptide having the sequence of aminoacid residues from 1 or about 19 to about 197, inclusive of FIG. 6 (SEQID NO:11), or (b) the complement of the DNA molecule of (a).

[0102] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1122 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 104 andabout 640, inclusive, of FIG. 5 (SEQ ID NO:10). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0103] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203552 (DNA62377-1381), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203552 (DNA62377-1381).

[0104] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 19 to about 197, inclusive of FIG. 6 (SEQID NO:11), or the complement of the DNA of (a).

[0105] In a further aspect, the invention concerns an isolated nucleicacid molecule produced by hybridizing a test DNA molecule understringent conditions with (a) a DNA molecule encoding a PRO1122polypeptide having the sequence of amino acid residues from about 19 toabout 197, inclusive of FIG. 6 (SEQ ID NO:11), or (b) the complement ofthe DNA molecule of (a), and, if the DNA molecule has at least about an80% sequence identity, preferably at least about an 85% sequenceidentity, more preferably at least about a 90% sequence identity, mostpreferably at least about a 95% sequence identity to (a) or (b),isolating the test DNA molecule.

[0106] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 19to about 197, inclusive of FIG. 6 (SEQ ID NO:11), or (b) the complementof the DNA of (a).

[0107] In another embodiment, the invention provides isolated PRO1122polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0108] In a specific aspect, the invention provides isolated nativesequence PRO1122 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 19 through 197 of FIG. 6 (SEQ IDNO:11).

[0109] In another aspect, the invention concerns an isolated PRO1122polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues19 to about 197, inclusive of FIG. 6 (SEQ ID NO:11).

[0110] In a further aspect, the invention concerns an isolated PRO1122polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 19through 197 of FIG. 6 (SEQ ID NO:11).

[0111] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1122 polypeptide havingthe sequence of amino acid residues from about 19 to about 197,inclusive of FIG. 6 (SEQ ID NO:1), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0112] 4. PRO1889

[0113] A cDNA clone (DNA77623-2524) has been identified, having homologyto nucleic acid encoding E48 antigen protein, that encodes a novelpolypeptide, designated in the present application as “PRO1889”.

[0114] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1889 polypeptide In oneaspect, the isolated nucleic acid comprises DNA having at least about80% sequence identity, preferably at least about 85% sequence identity,more preferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding aPRO1889 polypeptide having the sequence of amino acid residues fromabout 1 or about 21 to about 97, inclusive of FIG. 8 (SEQ ID NO:16), or(b) the complement of the DNA molecule of (a).

[0115] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1889 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about nucleotides 39 orabout 99 and about: 329, inclusive, of FIG. 7 (SEQ ID NO:15).Preferably, hybridization occurs under stringent hybridization and washconditions.

[0116] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203546 (DNA77623-2524) or (b) the complement of the nucleicacid molecule of (a). In a preferred embodiment, the nucleic acidcomprises a DNA encoding the same mature polypeptide encoded by thehuman protein cDNA in ATCC Deposit No. 203546 (DNA77623-2524).

[0117] In still a further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues 1 or about 21 to about 97, inclusive of FIG. 8 (SEQID NO:16), or (b) the complement of the DNA of (a).

[0118] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least 315 nucleotides and produced byhybridizing a test DNA molecule under stringent conditions with (a) aDNA molecule encoding a PRO1889 polypeptide having the sequence of aminoacid residues from 1 or about 21 to about 97, inclusive of FIG. 8 (SEQID NO:16), or (b) the complement of the DNA molecule of (a), and, if theDNA molecule has at least about an 80% sequence identity, prefereably atleast about an 85% sequence identity, more preferably at least about a90% sequence identity, most preferably at least about a 95% sequenceidentity to (a) or (b), isolating the test DNA molecule.

[0119] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1889 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,or is complementary to such encoding nucleic acid molecule. The signalpeptide has been tentatively identified as extending from about aminoacid position 1 to about amino acid position 20 in the sequence of FIG.8 (SEQ ID NO:16).

[0120] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 1or about 21 to about 97, inclusive of FIG. 8 (SEQ ID NO:16), or (b) thecomplement of the DNA of (a).

[0121] Another embodiment is directed to fragments of a PRO1889polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 to about 80 nucleotides inlength, preferably from about 20 to about 60 nucleotides in length, morepreferably from about 20 to about 50 nucleotides in length and mostpreferably from about 20 to about 40 nucleotides in length and may bederived from the nucleotide sequence shown in FIG. 7 (SEQ ID NO:15).

[0122] In another embodiment, the invention provides isolated PRO1889polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0123] In a specific aspect, the invention provides isolated nativesequence PRO1889 polypeptide, which in certain embodiments, includes anamino acid sequence comprising residues 1 or about 21 to about 97 ofFIG. 8 (SEQ ID NO:16).

[0124] In another aspect, the invention concerns an isolated PRO1889polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues1 or about 21 to about 97, inclusive of FIG. 8 (SEQ ID NO:16).

[0125] In a further aspect, the invention concerns an isolated PRO1889polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 1 orabout 21 to about 97, inclusive of FIG. 8 (SEQ ID NO:16).

[0126] In yet another aspect, the invention concerns an isolated PRO1889polypeptide, comprising the sequence of amino acid residues 0.1 or about21 to about 97, inclusive of FIG. 8 (SEQ ID NO:16), or a fragmentthereof sufficient to provide a binding site for an anti-PRO1889antibody. Preferably, the PRO1889 fragment retains a qualitativebiological activity of a native PRO1889 polypeptide.

[0127] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1889 polypeptide havingthe sequence of amino acid residues from about 1 or about 21 to about97, inclusive of FIG. 8 (SEQ ID NO:16), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0128] In another embodiment, the invention concerns a method fordetermining the presence of a PRO1889 polypeptide comprising exposing acell suspected of containing the polypeptide to an anti-PRO1889 antibodyand determining binding of the antibody to the cell.

[0129] In yet another embodiment, the present invention relates to amethod of diagnosing the presence of a cancerous cell in a mammal,comprising detecting the level of expression of a gene encoding aPRO1889 polypeptide (a) in a test sample of tissue cells obtained fromthe mammal, and (b) in a control sample of known normal tissue cells ofthe same cell type, wherein a higher expression level in the test sampleindicates the presence of a cancerous cell, particularly a canceroussquamous cell, in the mammal.

[0130] A further embodiment is a method for identifying a compoundcapable of inhibiting the expression and/or activity of a PRO1889polypeptide by contacting a candidate compound with a PRO1889polypeptide under conditions and for time sufficient to allow these twocompounds to interact. In a specific aspect, either the candidatecompound or the PRO1889 polypeptide is immobilized on a solid support.In another aspect, the non-immobilized component carries a detectablelabel.

[0131] In a further embodiment, the present invention provides a methodof diagnosing tumor in a mammal, comprising detecting the level ofexpression of a gene encoding a PRO1889 polypeptide (a) in a test sampleof tissue cells obtained from the mammal, and (b) in a control sample ofknown normal tissue cells of the same cell type, wherein a higherexpression level in the test sample indicates the presence of tumor inthe mammal from which the test tissue cells were obtained.

[0132] In another embodiment, the present invention provides a method ofdiagnosing tumor in a mammal, comprising (a) contacting an anti-PRO1889antibody with a test sample of the tissue cells obtained from themammal, and (b) detecting the formation of a complex between theanti-PRO1889 and the PRO1889 polypeptide in the test sample. Thedetection may be qualitative or quantitative, and may be performed incomparison with monitoring the complex formation in a control sample ofknown normal tissue cells of the same cell type. A larger quantity ofcomplexes formed in the test sample indicates the presence of tumor inthe mammal from which the test tissue cells were obtained. The antibodypreferably carries a detectable label. Complex formation can bemonitored, for example, by light microscopy, flow cytometry,fluorimetry, or other techniques known in the art. Preferably, the testsample is obtained from an individual mammal suspected to haveneoplastic cell growth or proliferation (e.g., cancerous cells).

[0133] In another embodiment, the present invention provides a cancerdiagnostic kit, comprising an anti-PRO1889 antibody and a carrier (e.g.a buffer) in suitable packaging. The kit preferably containsinstructions for using the antibody to detect the PRO1889 polypeptide.

[0134] In yet another embodiment, the invention provides a method forinhibiting the growth of tumor cells comprising exposing a cell whichoverexpresses a PRO1889 polypeptide to an effective amount of an agentinhibiting the expression and/or activity of the PRO1889 polypeptide.The agent preferably is an anti-PRO1889 polypeptide, a small organic andinorganic peptide, phosphopeptide, antisense or ribozyme molecule, or atriple helix molecule. In a specific aspect, the agent, e.g.,anti-PRO1889 antibody induces cell death. In a further aspect, the tumorcells are further exposed to radiation treatment and/or a cytotoxic orchemotherapeutic agent.

[0135] In a further embodiment, the invention concerns an article ofmanufacture, comprising:

[0136] a container;

[0137] a label on the container, and

[0138] a composition comprising an active agent contained within thecontainer; wherein the composition is effective for inhibiting thegrowth of tumor cells, the label on the container indicates that thecomposition can be used for treating conditions characterized byoverexpression of a PRO1889 polypeptide, and the active agent in thecomposition is an agent inhibiting the expression and/or activity of thePRO1889 polypeptide. In a preferred aspect, the active agent is ananti-PRO1889 antibody.

[0139] 5. PRO1890

[0140] A cDNA clone (DNA79230-2525) has been identified, having homologyto nucleic acid encoding a lectin protein that encodes a novelpolypeptide, designated in the present application as “PRO1890”.

[0141] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1890 polypeptide.

[0142] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO1890 polypeptide having the sequence of aminoacid residues from about 1 or about 22 to about 273, inclusive of FIG.10 (SEQ ID NO:18), or (b) the complement of the DNA molecule of (a).

[0143] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1890 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about nucleotides 378 orabout 441 and about 1196, inclusive, of FIG. 9 (SEQ ID NO:17).Preferably, hybridization occurs under stringent hybridization and washconditions.

[0144] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203549 (DNA79230-2525) or (b) the complement of the nucleicacid molecule of (a). In a preferred embodiment, the nucleic acidcomprises a DNA encoding the same mature polypeptide encoded by thehuman protein cDNA in ATCC Deposit No. 203549 (DNA79230-2525).

[0145] In still a further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues 1 or about 22 to about 273, inclusive of FIG. 10(SEQ ID NO:18), or (b) the complement of the DNA of (a).

[0146] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least 475 nucleotides and produced byhybridizing a test DNA molecule under stringent conditions with (a) aDNA molecule encoding a PRO1890 polypeptide having the sequence of aminoacid residues from 1 or about 22 to about 273, inclusive of FIG. 10 (SEQID NO:18), or (b) the complement of the DNA molecule of (a), and, if theDNA molecule has at least about an 80% sequence identity, prefereably atleast about an 85% sequence identity, more preferably at least about a90% sequence identity, most preferably at least about a 95% sequenceidentity to (a) or (b), isolating the test DNA molecule.

[0147] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1890 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble, i.e., transmembrane domain deleted or inactivatedvariants, or is complementary to such encoding nucleic acid molecule.The signal peptide has been tentatively identified as extending fromabout amino acid position 1 to about amino acid position 21 in thesequence of FIG. 10 (SEQ ID NO:18). The transmembrane domain has beententatively identified as extending from about amino acid position 214to about amino acid position 235 in the PRO1890 amino acid sequence(FIG. 10, SEQ ID NO:18).

[0148] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 1or about 22 to about 273, inclusive of FIG. 10 (SEQ ID NO:18), or (b)the complement of the DNA of (a).

[0149] Another embodiment is directed to fragments of a PRO1890polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 to about 80 nucleotides inlength, preferably from about 20 to about 60 nucleotides in length, morepreferably from about 20 to about 50 nucleotides in length and mostpreferably from about 20 to about 40 nucleotides in length and may bederived from the nucleotide sequence shown in FIG. 9 (SEQ ID NO:17).

[0150] In another embodiment, the invention provides isolated PRO1890polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0151] In a specific aspect, the invention provides isolated nativesequence PRO1890 polypeptide, which in certain embodiments, includes anamino acid sequence comprising residues 1 or about 22 to about 273 ofFIG. 10 (SEQ ID NO:18).

[0152] In another aspect, the invention concerns an isolated PRO1890polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues1 or about 22 to about 273, inclusive of FIG. 10 (SEQ ID NO:18).

[0153] In a further aspect, the invention concerns an isolated PRO1890polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 1 orabout 22 to about 273, inclusive of FIG. 10 (SEQ ID NO:18).

[0154] In yet another aspect, the invention concerns an isolated PRO1890polypeptide, comprising the sequence of amino acid residues 1 or about22 to about 273, inclusive of FIG. 10 (SEQ ID NO:18), or a fragmentthereof sufficient to provide a binding site for an anti-PRO1890antibody. Preferably, the PRO1890 fragment retains a qualitativebiological activity of a native PRO1890 polypeptide.

[0155] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1890 polypeptide havingthe sequence of amino acid residues from about 1 or about 22 to about273, inclusive of FIG. 10 (SEQ ID NO:18), or (b) the complement of theDNA molecule of (a), and if the test DNA molecule has at least about an80% sequence identity, preferably at least about an 85% sequenceidentity, more preferably at least about a 90% sequence identity, mostpreferably at least about a 95% sequence identity to (a) or (b), (ii)culturing a host cell comprising the test DNA molecule under conditionssuitable for expression of the polypeptide, and (iii) recovering thepolypeptide from the cell culture.

[0156] 6. PRO1887

[0157] A cDNA clone (DNA79862-2522) has been identified that encodes anovel polypeptide having homology to carboxylesterases and designated inthe present application as “PRO1887”.

[0158] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1887 polypeptide.

[0159] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO1887 polypeptide having the sequence of aminoacid residues from 1 or about 28 to about 571, inclusive of FIG. 12 (SEQID NO:23), or (b) the complement of the DNA molecule of (a).

[0160] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1887 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 87 andabout 1718, inclusive, of FIG. 11 (SEQ ID NO:22). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0161] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203550 (DNA79862-2522), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203550 (DNA79862-2522).

[0162] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 28 to about 571, inclusive of FIG. 12(SEQ ID NO:23), or the complement of the DNA of (a).

[0163] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO1887 polypeptide having the sequence of amino acid residues fromabout 28 to about 571, inclusive of FIG. 12 (SEQ ID NO:23), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0164] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1887 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position27 in the sequence of FIG. 12 (SEQ ID NO:23). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 226 to about amino acid position 245 in the PRO1887 amino acidsequence (FIG. 12, SEQ ID NO:23).

[0165] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 28to about 571, inclusive of FIG. 12 (SEQ ID NO:23), or (b) the complementof the DNA of (a).

[0166] Another embodiment is directed to fragments of a PRO1887polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 to about 80 nucleotides inlength, preferably from about 20 to about 60 nucleotides in length, morepreferably from about 20 to about 50 nucleotides in length, and mostpreferably from about 20 to about 40 nucleotides in length.

[0167] In another embodiment, the invention provides isolated PRO1887polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0168] In a specific aspect, the invention provides isolated nativesequence PRO1887 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 28 to 571 of FIG. 12 (SEQ ID NO:23).

[0169] In another aspect, the invention concerns an isolated PRO1887polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues28 to about 571, inclusive of FIG. 12 (SEQ ID NO:23).

[0170] In a further aspect, the invention concerns an isolated PRO1887polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 28 to571 of FIG. 12 (SEQ ID NO:23).

[0171] In yet another aspect, the invention concerns an isolated PRO1887polypeptide, comprising the sequence of amino acid residues 28 to about571, inclusive of FIG. 12 (SEQ ID NO:23), or a fragment thereofsufficient to provide a binding site for an anti-PRO1887 antibody.Preferably, the PRO1887 fragment retains a qualitative biologicalactivity of a native PRO1887 polypeptide.

[0172] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1887 polypeptide havingthe sequence of amino acid residues from about 28 to about 571,inclusive of FIG. 12 (SEQ ID NO:23), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0173] 7. PRO1785

[0174] A cDNA clone (DNA80136-2503) has been identified that encodes anovel polypeptide having sequence identity with peroxidases anddesignated in the present application as “PRO1785.” In one embodiment,the invention provides an isolated nucleic acid molecule comprising DNAencoding a PRO1785 polypeptide.

[0175] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least abo at 95% sequence identity to (a) a DNAmolecule encoding a PRO1785 polypeptide having the sequence of aminoacid residues from 1 or about 32 to about 209, inclusive of FIG. 14 (SEQID NO:29), or (b) the complement of the DNA molecule of (a).

[0176] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO1785 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 95 andabout 628, inclusive., of FIG. 13 (SEQ ID NO:28). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0177] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203541 (DNA80136-2503), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203541 (DNA80136-2503).

[0178] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 32 to about 209, inclusive of FIG. 14(SEQ ID NO:29), or the complement of the DNA of (a).

[0179] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO1785 polypeptide having the sequence of amino acid residues fromabout 32 to about 209, inclusive of FIG. 14 (SEQ ID NO:29), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0180] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO1785 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble, i.e. transmembrane domain deleted or inactivatedvariants, or is complementary to such encoding nucleic acid molecule.The signal peptide has been tentatively identified as extending fromamino acid position 1 through about amino acid position 31 in thesequence of FIG. 14 (SEQ ID NO:29). The transmembrane domain has beententatively identified as extending from about amino acid position 18through about amino acid position 37 in the PRO1785 amino acid sequence(FIG. 14, SEQ ID NO:29).

[0181] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 32to about 209, inclusive of FIG. 14 (SEQ ID NO:29), or (b) the complementof the DNA of (a).

[0182] Another embodiment is directed to fragments of a PRO1785polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 to about 80 nucleotides inlength, preferably from about 20 to about 60 nucleotides in length, morepreferably from about 20 to about 50 nucleotides in length, and mostpreferably from about 20 to about 40 nucleotides in length.

[0183] In another embodiment, the invention provides isolated PRO1785polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0184] In a specific aspect, the invention provides isolated nativesequence PRO1785 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 32 through 209 of FIG. 14 (SEQ IDNO:29).

[0185] In another aspect, the invention concerns an isolated PRO1785polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues32 to about 209, inclusive of FIG. 14 (SEQ ID NO:29).

[0186] In a further aspect, the invention concerns an isolated PRO1785polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 32through 209 of FIG. 14 (SEQ ID NO:29).

[0187] In yet another aspect, the invention concerns an isolated PRO1785polypeptide, comprising the sequence of amino acid residues 32 to about209, inclusive of FIG. 14 (SEQ ID NO:29), or a fragment thereofsufficient to provide a binding site for an anti-PRO1785 antibody.Preferably, the PRO1785 fragment retains a qualitative biologicalactivity of a native PRO1785 polypeptide.

[0188] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO1785 polypeptide havingthe sequence of amino acid residues from about 32 to about 209,inclusive of FIG. 14 (SEQ ID NO:29), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0189] 8. PRO4353

[0190] A cDNA clone (DNA80145-2594) has been identified that encodes anovel polypeptide having homology to semaphorin Z and designated in thepresent application as “PRO4353”.

[0191] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4353 polypeptide.

[0192] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4353 polypeptide having the sequence of aminoacid residues from 1 or about 26 to about 888, inclusive of FIG. 16 (SEQID NO:35), or (b) the complement of the DNA molecule of (a).

[0193] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4353 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 94 andabout 2682, inclusive, of FIG. 15 (SEQ ID NO:34). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0194] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 204-PTA (DNA80145-2594), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 204-PTA (DNA80145-2594).

[0195] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 26 to about 888, inclusive of FIG. 16(SEQ ID NO:35), or the complement of the DNA of (a).

[0196] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4353 polypeptide having the sequence of amino acid residues fromabout 26 to about 888, inclusive of FIG. 16 (SEQ ID NO:35), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0197] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4353 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position25 in the sequence of FIG. 16 (SEQ ID NO:35). The transmembrane domainshave been tentatively identified as extending from about amino acidpositions 318-339 and 598-617 (FIG. 16, SEQ ID NO:35).

[0198] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 26to about 888, inclusive of FIG. 16 (SEQ ID NO:35), or (b) the complementof the DNA of (a).

[0199] Another embodiment is directed to fragments of a PRO4353polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0200] In another embodiment, the invention provides isolated PRO4353polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0201] In a specific aspect, the invention provides isolated nativesequence PRO4353 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 26 through 888 of FIG. 16 (SEQ IDNO:35).

[0202] In another aspect, the invention concerns an isolated PRO4353polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues26 to about 888, inclusive of FIG. 16 (SEQ ID NO:35).

[0203] In a further aspect, the invention concerns an isolated PRO4353polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 26through 888 of FIG. 16 (SEQ ID NO:35).

[0204] In yet another aspect, the invention concerns an isolated PRO4353polypeptide, comprising the sequence of amino acid residues 26 to about888, inclusive of FIG. 16 (SEQ ID NO:35), or a fragment thereofsufficient to provide a binding site for an anti-PRO4353 antibody.Preferably, the PRO4353 fragment retains a qualitative biologicalactivity of a native PRO4353 polypeptide.

[0205] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4353 polypeptide havingthe sequence of amino acid residues from about 26 to about 888,inclusive of FIG. 16 (SEQ ID NO:35), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0206] 9. PRO4357

[0207] A cDNA clone (DNA84917-2597) has been identified that encodes anovel polypeptide having homology to “BK158_(—)1” and designated in thepresent application as “PRO4357”.

[0208] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4357 polypeptide.

[0209] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4357 polypeptide having the sequence of aminoacid residues from 1 or about 18 to about 502, inclusive of FIG. 18 (SEQID NO:40), or (b) the complement of the DNA molecule of (a).

[0210] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4357 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 337 andabout 1791, inclusive, of FIG. 17 (SEQ ID NO:39). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0211] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203863 (DNA84917-2597), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203863 (DNA84917-2597).

[0212] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 18 to about 502, inclusive of FIG. 18(SEQ ID NO:40), or the complement of the DNA of (a).

[0213] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4357 polypeptide having the sequence of amino acid residues fromabout 18 to about 502, inclusive of FIG. 18 (SEQ ID NO:40), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0214] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 18to about 502, inclusive of FIG. 18 (SEQ ID NO:40), or (b) the complementof the DNA of (a).

[0215] Another embodiment is directed to fragments of a PRO4357polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0216] In another embodiment, the invention provides isolated PRO4357polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0217] In a specific aspect, the invention provides isolated nativesequence PRO4357 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 18 through 502 of FIG. 18 (SEQ IDNO:40).

[0218] In another aspect, the invention concerns an isolated PRO4357polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues18 to about 502, inclusive of FIG. 18 (SEQ ID NO:40).

[0219] In a further aspect, the invention concerns an isolated PRO4357polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 18through 502 of FIG. 18 (SEQ ID NO:40).

[0220] In yet another aspect, the invention concerns an isolated PRO4357polypeptide, comprising the sequence of amino acid residues 18 to about502, inclusive of FIG. 18 (SEQ ID NO:40), or a fragment thereofsufficient to provide a binding site for an anti-PRO4357 antibody.Preferably, the PRO4357 fragment retains a qualitative biologicalactivity of a native PRO4357 polypeptide.

[0221] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4357 polypeptide havingthe sequence of amino acid residues from about 18 to about 502,inclusive of FIG. 18 (SEQ ID NO:40), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0222] 10. PRO4405

[0223] A cDNA clone (DNA84920-2614) has been identified that encodes anovel polypeptide designated in the present application as “PRO4405”.

[0224] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4405 polypeptide.

[0225] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4405 polypeptide having the sequence of aminoacid residues from 1 or about 35 to about 310, inclusive of FIG. 20 (SEQID NO:45), or (b) the complement of the DNA molecule of (a).

[0226] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4405 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 181 andabout 1008, inclusive, of FIG. 19 (SEQ ID NO:44). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0227] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203966 (DNA84920-2614), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203966 (DNA84920-2614).

[0228] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 35 to about 310, inclusive of FIG. 20(SEQ ID NO:45), or the complement of the DNA of (a).

[0229] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4405 polypeptide having the sequence of amino acid residues fromabout 35 to about 310, inclusive of FIG. 20 (SEQ ID NO:45), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0230] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4405 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position34 in the sequence of FIG. 20 (SEQ ID NO:45). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 58 through about amino acid position 76 in the PRO4405 aminoacid sequence (FIG. 20, SEQ ID NO:45) and may be a type II transmembranedomain.

[0231] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 35to about 310, inclusive of FIG. 20 (SEQ ID NO:45), or (b) the complementof the DNA of (a).

[0232] Another embodiment is directed to fragments of a PRO4405polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0233] In another embodiment, the invention provides isolated PRO4405polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0234] In a specific aspect, the invention provides isolated nativesequence PRO4405 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 35 through 310 of FIG. 20 (SEQ IDNO:45).

[0235] In another aspect, the invention concerns an isolated PRO4405polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues35 to about 310, inclusive of FIG. 20 (SEQ ID NO:45).

[0236] In a further aspect, the invention concerns an isolated PRO4405polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 35through 310 of FIG. 20 (SEQ ID NO:45).

[0237] In yet another aspect, the invention concerns an isolated PRO4405polypeptide, comprising the sequence of amino acid residues 35 to about310, inclusive of FIG. 20 (SEQ ID NO:45), or a fragment thereofsufficient to provide a binding site for an anti-PRO4405 antibody.Preferably, the PRO4405 fragment retains a qualitative biologicalactivity of a native PRO4405 polypeptide.

[0238] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4405 polypeptide havingthe sequence of amino acid residues from about 35 to about 310,inclusive of FIG. 20 (SEQ ID NO:45), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0239] 11. PRO4356

[0240] A cDNA clone (DNA86576-2595) has been identified that encodes anovel polypeptide having homology to MAGPIAP and designated in thepresent application as “PRO4356”.

[0241] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4356 polypeptide.

[0242] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about at 95% sequence identity to (a) a DNAmolecule encoding a PRO4356 polypeptide having the sequence of aminoacid residues from 1 or about 20 to about 251, inclusive of FIG. 22 (SEQID NO:50), or (b) the complement of the DNA molecule of (a).

[0243] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4356 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 112 andabout 807, inclusive, of FIG. 21 (SEQ ID NO:49). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0244] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203868 (DNA86576-2595), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203868 (DNA86576-2595).

[0245] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypepticle havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 20 to about 251, inclusive of FIG. 22(SEQ ID NO:50), or the complement of the DNA of (a).

[0246] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4356 polypeptide having the sequence of amino acid residues fromabout 20 to about 251, inclusive of FIG. 22 (SEQ ID NO:50), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0247] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4356 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position19 in the sequence of FIG. 22 (SEQ ID NO:50). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 233 through about amino acid position 251 in the PRO4356 aminoacid sequence (FIG. 22, SEQ ID NO:50).

[0248] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 20to about 251, inclusive of FIG. 22 (SEQ ID NO:50), or (b) the complementof the DNA of (a).

[0249] Another embodiment is directed to fragments of a PRO4356polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0250] In another embodiment, the invention provides isolated PRO4356polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0251] In a specific aspect, the invention provides isolated nativesequence PRO4356 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 20 through 251 of FIG. 22 (SEQ IDNO:50).

[0252] In another aspect, the invention concerns an isolated PRO4356polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues20 to about 251, inclusive of FIG. 22 (SEQ ID NO:50).

[0253] In a further aspect, the invention concerns an isolated PRO4356polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 20through 251 of FIG. 22 (SEQ ID NO:50).

[0254] In yet another aspect, the invention concerns an isolated PRO4356polypeptide, comprising the sequence of amino acid residues 20 to about251, inclusive of FIG. 22 (SEQ ID NO:50), or a fragment thereofsufficient to provide a binding site for an anti-PRO4356 antibody.Preferably, the PRO4356 fragment retains a qualitative biologicalactivity of a native PRO4356 polypeptide.

[0255] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4356 polypeptide havingthe sequence of amino acid residues from about 20 to about 251,inclusive of FIG. 22 (SEQ ID NO:50), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0256] 12. PRO4352

[0257] A cDNA clone (DNA87976-2593) has been identified that encodes anovel polypeptide having homology to protocadherin pc3 designated in thepresent application as “PRO4352”.

[0258] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4352 polypeptide.

[0259] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4352 polypeptide having the sequence of aminoacid residues from 1 or about 27 to about 800, inclusive of FIG. 24 (SEQID NO:52), or (b) the complement of the DNA molecule of (a).

[0260] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4352 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 257 andabout 2578, inclusive, of FIG. 23 (SEQ ID NO:51). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0261] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203888 (DNA87976-2593), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNAK encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203888 (DNA87976-2593).

[0262] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 27 to about 800, inclusive of FIG. 24(SEQ ID NO:52), or the complement of the DNA of (a).

[0263] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4352 polypeptide having the sequence of amino acid residues fromabout 27 to about 800, inclusive of FIG. 24 (SEQ ID NO:52), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0264] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4352 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule, The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position26 in the sequence of FIG. 24 (SEQ ID NO:52). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 687 through about amino acid position 711 in the PRO4352 aminoacid sequence (FIG. 24, SEQ ID NO:52).

[0265] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 27to about 800, inclusive of FIG. 24 (SEQ ID NO:52), or (b) the complementof the DNA of (a).

[0266] Another embodiment is directed to fragments of a PRO4352polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0267] In another embodiment, the invention provides isolated PRO4352polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0268] In a specific aspect, the invention provides isolated nativesequence PRO4352 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 27 through 800 of FIG. 24 (SEQ IDNO:52).

[0269] In another aspect, the invention concerns an isolated PRO4352polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues27 to about 800, inclusive of FIG. 24 (SEQ ID NO:52).

[0270] In a further aspect, the invention concerns an isolated PRO4352polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 27through 800 of FIG. 24 (SEQ ID NO:52).

[0271] In yet another aspect, the invention concerns an isolated PRO4352polypeptide, comprising the sequence of amino acid residues 27 to about800, inclusive of FIG. 24 (SEQ ID NO:52), or a fragment thereofsufficient to provide a binding site for an anti-PRO4352 antibody.Preferably, the PRO4352 fragment retains a qualitative biologicalactivity of a native PRO4352 polypeptide.

[0272] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4352 polypeptide havingthe sequence of amino acid residues from about 27 to about 800,inclusive of FIG. 24 (SEQ ID NO:52), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0273] 13. PRO4380

[0274] A cDNA clone (DNA92234-2602) has been identified that encodes anovel polypeptide designated in the present application as “PRO4380”.

[0275] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4380 polypeptide.

[0276] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4380 polypeptide having the sequence of aminoacid residues from 1 or about 27 to about 507, inclusive of FIG. 26 (SEQID NO:57), or (b) the complement of the DNA molecule of (a).

[0277] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4380 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 279 andabout 1721, inclusive, of FIG. 25 (SEQ ID NO:56). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0278] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203948 (DNA92234-2602), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203948 (DNA92234-2602).

[0279] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 27 to about 507, inclusive of FIG. 26(SEQ ID NO:57), or the complement of the DNA of (a).

[0280] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4380 polypeptide having the sequence of amino acid residues fromabout 27 to about 507, inclusive of FIG. 26 (SEQ ID NO:57), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0281] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4380 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position I through about amino acid position26 in the sequence of FIG. 26 (SEQ ID NO:57). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 273 through about amino acid position 292 in the PRO4380 aminoacid sequence (FIG. 26, SEQ ID NO:57).

[0282] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 27to about 507, inclusive of FIG. 26 (SEQ ID NO:57), or (b) the complementof the DNA of (a).

[0283] Another embodiment is directed to fragments of a PRO4380polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0284] In another embodiment, the invention provides isolated PRO4380polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0285] In a specific aspect, the invention provides isolated nativesequence PRO4380 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 27 through 507 of FIG. 26 (SEQ IDNO:57).

[0286] In another aspect, the invention concerns an isolated PRO4380polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues27 to about 507, inclusive of FIG. 26 (SEQ ID NO:57).

[0287] In a further aspect, the invention concerns an isolated PRO4380polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 27through 507 of FIG. 26 (SEQ ID NO:57).

[0288] In yet another aspect, the invention concerns an isolated PRO4380polypeptide, comprising the sequence of amino acid residues 27 to about507, inclusive of FIG. 26 (SEQ ID NO:57), or a fragment thereofsufficient to provide a binding site for an anti-PRO4380 antibody.Preferably, the PRO4380 fragment retains a qualitative biologicalactivity of a native PRO4380 polypeptide.

[0289] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4380 polypeptide havingthe sequence of amino acid residues from about 27 to about 507,inclusive of FIG. 26 (SEQ ID NO:57), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0290] 14. PRO4354

[0291] A cDNA clone (DNA92256-2596) has been identified that encodes anovel polypeptide designated in the present application as “PRO4354”.

[0292] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4354 polypeptide.

[0293] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4354 polypeptide having the sequence of aminoacid residues from about 22 to about 248, inclusive of FIG. 28 (SEQ IDNO:59), or (b) the complement of the DNA molecule of (a).

[0294] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4354 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 171 andabout 851, inclusive, of FIG. 27 (SEQ ID NO:58). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0295] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203891 (DNA92256-2596), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203891 (DNA92256-2596).

[0296] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 22 to about 248, inclusive of FIG. 28(SEQ ID NO:59), or the complement of the DNA of (a).

[0297] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4354 polypeptide having the sequence of amino acid residues fromabout 22 to about 248, inclusive of FIG. 28 (SEQ ID NO:59), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0298] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 22to about 248, inclusive of FIG. 28 (SEQ ID NO:59), or (b) the complementof the DNA of (a).

[0299] Another embodiment is directed to fragments of a PRO4354polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0300] In another embodiment, the invention provides isolated PRO4354polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0301] In a specific aspect, the invention provides isolated nativesequence PRO4354 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 22 through 248 of FIG. 28 (SEQ IDNO:59).

[0302] In another aspect, the invention concerns an isolated PRO4354polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues22 to about 248, inclusive of FIG. 28 (SEQ ID NO:59).

[0303] In a further aspect, the invention concerns an isolated PRO4354polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 22through 248 of FIG. 28 (SEQ ID NO:59).

[0304] In yet another aspect, the invention concerns an isolated PRO4354polypeptide, comprising the sequence of amino acid residues 22 to about248, inclusive of FIG. 28 (SEQ ID NO:59), or a fragment thereofsufficient to provide a binding site for an anti-PRO4354 antibody.Preferably, the PRO4354 fragment retains a qualitative biologicalactivity of a native PRO4354 polypeptide.

[0305] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4354 polypeptide havingthe sequence of amino acid residues from about 22 to about 248,inclusive of FIG. 28 (SEQ ID NO:59), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0306] 15. PRO4408

[0307] A cDNA clone (DNA92274-2617) has been identified that encodes anovel polypeptide designated in the present application as “PRO4408”.

[0308] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4408 polypeptide.

[0309] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4408 polypeptide having the sequence of aminoacid residues from about 23 to about 223, inclusive of FIG. 30 (SEQ IDNO:61), or (b) the complement of the DNA molecule of (a).

[0310] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4408 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 155 andabout 757, inclusive, of FIG. 29 (SEQ ID NO:60). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0311] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203971 (DNA92274-2617), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNAK encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203971 (DNA92274-2617).

[0312] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 23 to about 223, inclusive of FIG. 30(SEQ ID NO:61), or the complement of the DNA of (a).

[0313] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4408 polypeptide having the sequence of amino acid residues fromabout 23 to about 223, inclusive of FIG. 30 (SEQ ID NO:61), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0314] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 23to about 223, inclusive of FIG. 30 (SEQ ID NO:61), or (b) the complementof the DNA of (a).

[0315] Another embodiment is directed to fragments of a PRO4408polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0316] In another embodiment, the invention provides isolated PRO4408polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0317] In a specific aspect, the invention provides isolated nativesequence PRO4408 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 23 through 223 of FIG. 30 (SEQ IDNO:61).

[0318] In another aspect, the invention concerns an isolated PRO4408polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues23 to about 223, inclusive of FIG. 30 (SEQ ID NO:61).

[0319] In a further aspect, the invention concerns an isolated PRO4408polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 23through 223 of FIG. 30 (SEQ ID NO:61).

[0320] In yet another aspect, the invention concerns an isolated PRO4408polypeptide, comprising the sequence of amino acid residues 23 to about223, inclusive of FIG. 30 (SEQ ID NO:61), or a fragment thereofsufficient to provide a binding site for an anti-PRO4408 antibody.Preferably, the PRO4408 fragment retains a qualitative biologicalactivity of a native PRO4408 polypeptide.

[0321] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4408 polypeptide havingthe sequence of amino acid residues from about 23 to about 223,inclusive of FIG. 30 (SEQ ID NO:61), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0322] 16. PRO5737

[0323] A cDNA clone (DNA92929-2534) has been identified that encodes anovel polypeptide having homology to IL-1 and is designated in thepresent application as “PRO5737”.

[0324] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO5737 polypeptide.

[0325] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO5737 polypeptide having the sequence of aminoacid residues from 1 or about 18 to about 134, inclusive of FIG. 32 (SEQID NO:63), or (b) the complement of the DNA molecule of (a).

[0326] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO5737 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 96 or about147 and about 497, inclusive, of FIG. 31 (SEQ ID NO:62). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0327] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203586 (DNA92929-2534), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203586 (DNA92929-2534).

[0328] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 18 to about 134, inclusive of FIG. 32(SEQ ID NO:63), or the complement of the DNA of (a).

[0329] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO5737 polypeptide having the sequence of amino acid residues fromabout 18 to about 134, inclusive of FIG. 32 (SEQ ID NO:63), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0330] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 18to about 134, inclusive of FIG. 32 (SEQ ID NO:63), or (1)) thecomplement of the DNA of (a).

[0331] Another embodiment is directed to fragments of a PRO5737polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0332] In another embodiment, the invention provides isolated PRO5737polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0333] In a specific aspect, the invention provides isolated nativesequence PRO5737 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 18 through 134 of FIG. 32 (SEQ IDNO:63).

[0334] In another aspect, the invention concerns an isolated PRO5737polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues18 to about 134, inclusive of FIG. 32 (SEQ ID NO:63).

[0335] In a further aspect, the invention concerns an isolated PRO5737polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 18through 134 of FIG. 32 (SEQ ID NO:63).

[0336] In yet another aspect, the invention concerns an isolated PRO5737polypeptide, comprising the sequence of amino acid residues 18 to about134, inclusive of FIG. 32 (SEQ ID NO:63), or a fragment thereofsufficient to provide a binding site for an anti-PRO5737 antibody.Preferably, the PRO5737 fragment retains a qualitative biologicalactivity of a native PRO5737 polypeptide.

[0337] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO5737 polypeptide havingthe sequence of amino acid residues from about 18 to about 134,inclusive of FIG. 32 (SEQ ID NO:63), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0338] 17. PRO4425

[0339] A cDNA clone (DNA93011-2637) has been identified that encodes anovel polypeptide having homology to a protein in GenBank, accessionnumber HGS_RE295, and designated in the present application as“PRO4425”.

[0340] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4425 polypeptide.

[0341] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4425 polypeptide having the sequence of aminoacid residues from 1 OR about 20 to about 136, inclusive of FIG. 34 (SEQID NO:65), or (b) the complement of the DNA molecule of (a).

[0342] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4425 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 84 andabout 434, inclusive, of FIG. 33 (SEQ ID NO:64). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0343] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein CDNA in ATCCDeposit No. 20-PTA (DNA93011-2637), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 20-PTA (DNA93011-2637).

[0344] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 20 to about 136, inclusive of FIG. 34(SEQ ID NO:65), or the complement of the DNA of (a).

[0345] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4425 polypeptide having the sequence of amino acid residues fromabout 20 to about 136, inclusive of FIG. 34 (SEQ ID NO:65), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0346] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 20to about 136, inclusive of FIG. 34 (SEQ ID NO:65), or (b) the complementof the DNA of (a).

[0347] Another embodiment is directed to fragments of a PRO4425polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0348] In another embodiment, the invention provides isolated PRO4425polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0349] In a specific aspect, the invention provides isolated nativesequence PRO4425 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 20 through 136 of FIG. 34 (SEQ IDNO:65).

[0350] In another aspect, the invention concerns an isolated PRO4425polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues20 to about 136, inclusive of FIG. 34 (SEQ ID NO:65).

[0351] In a further aspect, the invention concerns an isolated PRO4425polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 20through 136 of FIG. 34 (SEQ ID NO:65).

[0352] In yet another aspect, the invention concerns an isolated PRO4425polypeptide, comprising the sequence of amino acid residues 20 to about136, inclusive of FIG. 34 (SEQ ID NO:65), or a fragment thereofsufficient to provide a binding site for an anti-PRO4425 antibody.Preferably, the PRO4425 fragment retains a qualitative biologicalactivity of a native PRO4425 polypeptide.

[0353] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4425 polypeptide havingthe sequence of amino acid residues from about 20 to about 136,inclusive of FIG. 34 (SEQ ID NO:65), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0354] 18. PRO5990

[0355] A cDNA clone (designated herein as DNA96042-2682) has beenidentified that has homology to nucleic acid encoding secretogranin andthat encodes a novel polypeptide, designated in the present applicationas “PRO5990”.

[0356] In one embodiment, the invention provides an isolated nucleicacid molecule comprising a nucleotide sequence that encodes a PRO5990polypeptide.

[0357] In one aspect, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculeencoding a PRO5990 polypeptide having the sequence of amino acidresidues from about 1 or about 22 to about 468, inclusive, of FIG. 36(SEQ ID NO:67), or (b) the complement of the DNA molecule of (a).

[0358] In another aspect, the isolated nucleic acid molecule comprises(a) a nucleotide sequence encoding a PRO5990 polypeptide having thesequence of amino acid residues from about 1 or about 22 to about 468,inclusive, of FIG. 36 (SEQ ID NO:67), or (b) the complement of thenucleotide sequence of (a).

[0359] In other aspects, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculehaving the sequence of nucleotides from about 265 or about 328 to about1668, inclusive, of FIG. 35 (SEQ ID NO:66), or (b) the complement of theDNA molecule of (a).

[0360] In another aspect, the isolated nucleic acid molecule comprises(a) the nucleotide sequence of from about 265 or about 328 to about1668, inclusive, of FIG. 35 (SEQ ID NO:66), or (b) the complement of thenucleotide sequence of (a).

[0361] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to (a) a DNAmolecule that encodes the same mature polypeptide encoded by the humanprotein cDNA deposited with the ATCC on Jul. 20, 1999 under ATCC DepositNo. 382-PTA (DNA96042-2682) or (b) the complement of the DNA molecule of(a). In a preferred embodiment, the isolated nucleic acid moleculecomprises (a) a nucleotide sequence encoding the same mature polypeptideencoded by the human protein cDNA deposited with the ATCC on Jul. 20,1999 under ATCC Deposit No. 382-PTA (DNA96042-2682) or (b) thecomplement of the nucleotide sequence of (a).

[0362] In another aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to (a) thefull-length polypeptide coding sequence of the human protein cDNAdeposited with the ATCC on Jul. 20, 1999 under ATCC Deposit No. 382-PTA(DNA96042-2682) or (b) the complement of the nucleotide sequence of (a).In a preferred embodiment, the isolated nucleic acid molecule comprises(a) the full-length polypeptide coding sequence of the DNA depositedwith the ATCC on Jul. 20, 1999 under ATCC Deposit No. 382-PTA(DNA96042-2682) or (b) the complement of the nucleotide sequence of (a).

[0363] In another aspect, the invention concerns an isolated nucleicacid molecule which encodes an active PRO5990 polypeptide as definedbelow comprising a nucleotide sequence that hybridizes to the complementof a nucleic acid sequence that encodes amino acids 1 or about 22 toabout 468, inclusive, of FIG. 36 (SEQ ID NO:67). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0364] In yet another aspect, the invention concerns an isolated nucleicacid molecule which encodes an active PRO5990 polypeptide as definedbelow comprising a nucleotide sequence that hybridizes to the complementof the nucleic acid sequence between about nucleotides 265 or about 328and about 1668, inclusive, of FIG. 35 (SEQ ID NO:66). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0365] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 1301 nucleotides and which isproduced by hybridizing a test DNA molecule under stringent conditionswith (a) a DNA molecule encoding a PRO5990 polypeptide having thesequence of amino acid residues from about 1 or about 22 to about 468,inclusive, of FIG. 36 (SEQ ID NO:67), or (b) the complement of the DNAmolecule of (a), and, if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 81% sequence identity,more preferably at least about an 82% sequence identity, yet morepreferably at least about an 83% sequence identity, yet more preferablyat least about an 84% sequence identity, yet more preferably at leastabout an 85% sequence identity, yet more preferably at least about an86% sequence identity, yet more preferably at least about an 87%sequence identity, yet more preferably at least about an 88% sequenceidentity, yet more preferably at least about an 89% sequence identity,yet more preferably at least about a 90% sequence identity, yet morepreferably at least about a 91% sequence identity, yet more preferablyat least about a 92% sequence identity, yet more preferably at leastabout a 93% sequence identity, yet more preferably at least a bout a 94%sequence identity, yet more preferably at least about a 95% sequenceidentity, yet more preferably at least about a 96% sequence identity,yet more preferably at least about a 97% sequence identity, yet morepreferably at least about a 98% sequence identity and yet morepreferably at least about a 99% sequence identity to (a) or (b), andisolating the test DNA molecule.

[0366] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) a nucleotide sequence encoding apolypeptide scoring at least about 80% positives, preferably at leastabout 81% positives, more preferably at least about 82% positives, yetmore preferably at least about 83% positives, yet more preferably atleast about 84% positives, yet more preferably at least about 85%positives, yet more preferably at least about 86% positives, yet morepreferably at least about 87% positives, yet more preferably at leastabout 88% positives, yet more preferably at least about 89% positives,yet more preferably at least about 90% positives, yet more preferably atleast about 91% positives, yet more preferably at least about 92%positives, yet more preferably at least about 93% positives, yet morepreferably at least about 94% positives, yet more preferably at leastabout 95% positives, yet more preferably at least about 96% positives,yet more preferably at least about 97% positives, yet more preferably atleast about 98% positives and yet more preferably at least about 99%positives when compared with the amino acid sequence of residues about 1or about 22 to 468, inclusive, of FIG. 36 (SEQ ID NO:67), or (b) thecomplement of the nucleotide sequence of (a).

[0367] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO5990 polypeptide without theN-terminal signal sequence and/or the initiating methionine, or iscomplementary to such encoding nucleic acid molecule. The signal peptidehas been tentatively identified as extending from about amino acidposition 1 to about amino acid position 21 in the sequence of FIG. 36(SEQ ID NO:67). It is noted, however, that the C-terminal boundary ofthe signal peptide may vary, but most likely by no more than about 5amino acids on either side of the signal peptide C-terminal boundary asinitially identified herein, wherein the C-terminal boundary of thesignal peptide may be identified pursuant to criteria routinely employedin the art for identifying that type of amino acid sequence element(e.g., Nielsen et al., Prot. Enz. 10:1-6 (1997) and von Heinje et al.,Nucl. Acids. Res. 14:4683-4690 (1986)). Moreover, it is also recognizedthat, in some cases, cleavage of a signal sequence from a secretedpolypeptide is not entirely uniform, resulting in more than one secretedspecies. These polypeptides, and the polynucleotides encoding them, arecontemplated by the present invention. As such, for purposes of thepresent application, the signal peptide of the PRO5990 polypeptide shownin FIG. 36 (SEQ ID NO:67) extends from amino acids 1 to X of FIG. 36(SEQ ID NO:67), wherein X is any amino acid from 16 to 26 of FIG. 36(SEQ ID NO:67). Therefore, mature forms of the PRO5990 polypeptide whichare encompassed by the present invention include those comprising aminoacids X to 468 of FIG. 36 (SEQ ID NO:67), wherein X is any amino acidfrom 16 to 26 of FIG. 36 (SEQ ID NO:67) and variants thereof asdescribed below. Isolated nucleic acid molecules encoding thesepolypeptides are also contemplated.

[0368] Another embodiment is directed to fragments of a PRO5990polypeptide coding sequence that may find use as, for example,hybridization probes or for encoding fragments of a PRO5990 polypeptidethat may optionally encode a polypeptide comprising a binding site foran anti-PRO5990 antibody. Such nucleic acid fragments are usually atleast about 20 nucleotides in length, preferably at least about 30nucleotides in length, more preferably at least about 40 nucleotides inlength, yet more preferably at least about 50 nucleotides in length, yetmore preferably at least about 60 nucleotides in length, yet morepreferably at least about 70 nucleotides in length, yet more preferablyat least about 80 nucleotides in length, yet more preferably at leastabout 90 nucleotides in length, yet more preferably at least about 100nucleotides in length, yet more preferably at least about 110nucleotides in length, yet more preferably at least about 120nucleotides in length, yet more preferably at least about 130nucleotides in length, yet more preferably at least about 140nucleotides in length, yet more preferably at least about 150nucleotides in length, yet more preferably at least about 160nucleotides in length, yet more preferably at least about 170nucleotides in length, yet more preferably at least about 180nucleotides in length, yet more preferably at least about 190nucleotides in length, yet more preferably at least about 200nucleotides in length, yet more preferably at least about 250nucleotides in length, yet more preferably at least about 300nucleotides in length, yet more preferably at least about 350nucleotides in length, yet more preferably at least about 400nucleotides in length, yet more preferably at least about 450nucleotides in length, yet more preferably at least about 500nucleotides in length, yet more preferably at least about 600nucleotides in length, yet more preferably at least about 700nucleotides in length, yet more preferably at least about 800nucleotides in length, yet more preferably at least about 900nucleotides in length and yet more preferably at least about 1000nucleotides in length, wherein in this context the term “about” meansthe referenced nucleotide sequence length plus or minus 10% of thatreferenced length. In a preferred embodiment, the nucleotide sequencefragment is derived from any coding region of the nucleotide sequenceshown in FIG. 35 (SEQ ID NO:66). It is noted that novel fragments of aPRO5990 polypeptide-encoding nucleotide sequence may be determined in aroutine manner by aligning the PRO5990 polypeptide-encoding nucleotidesequence with other known nucleotide sequences using any of a number ofwell known sequence alignment programs and determining which PRO5990polypeptide-encoding nucleotide sequence fragment(s) are novel. All ofsuch PRO5990 polypeptide-encoding nucleotide sequences are contemplatedherein and can be determined without undue experimentation. Alsocontemplated are the PRO5990 polypeptide fragments encoded by thesenucleotide molecule fragments, preferably those PRO5990 polypeptidefragments that comprise a binding site for an anti-PRO5990 antibody.

[0369] In another embodiment, the invention provides isolated PRO5990polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0370] In a specific aspect, the invention provides isolated nativesequence PRO5990 polypeptide, which in certain embodiments, includes anamino acid sequence comprising residues from about 1 or about 22 toabout 468 of FIG. 36 (SEQ ID NO:67).

[0371] In another aspect, the invention concerns an isolated PRO5990polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to the sequenceof amino acid residues from about 1 or about 22 to about 468, inclusive,of FIG. 36 (SEQ ID NO:67).

[0372] In a further aspect, the invention concerns an isolated PRO5990polypeptide comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to an aminoacid sequence encoded by the human protein CDNA deposited with the ATCCon Jul. 20, 1999 under ATCC Deposit No. 382-PTA (DNA96042-2682). In apreferred embodiment, the isolated PRO5990 polypeptide comprises anamino acid sequence encoded by the human protein cDNA deposited with theATCC on Jul. 20, 1999 under ATCC Deposit No. 382-PTA (DNA96042-2682).

[0373] In a further aspect, the invention concerns an isolated PRO5990polypeptide comprising an amino acid sequence scoring at least about 80%positives, preferably at least about 81% positives, more preferably atleast about 82% positives, yet more preferably at least about 83%positives, yet more preferably at least about 84% positives, yet morepreferably at least about 85% positives, yet more preferably at leastabout 86% positives, yet more preferably at least about 87% positives,yet more preferably at least about 88% positives, yet more preferably atleast about 89% positives, yet more preferably at least about 90%positives, yet more preferably at least about 91% positives, yet morepreferably at least about 92% positives, yet more preferably at leastabout 93% positives, yet more preferably at least about 94% positives,yet more preferably at least about 95% positives, yet more preferably atleast about 96% positives, yet more preferably at least about 97%positives, yet more preferably at least about 98% positives and yet morepreferably at least about 99% positives when compared with the aminoacid sequence of residues from about 1 or about 22 to about 468,inclusive, of FIG. 36 (SEQ ID NO:67).

[0374] In a specific aspect, the invention provides an isolated PRO5990polypeptide without the N-terminal signal sequence and/or the initiatingmethionine and is encoded by a nucleotide sequence that encodes such anamino acid sequence as hereinbefore described. Processes for producingthe same are also herein described, wherein those processes compriseculturing a host cell comprising a vector which comprises theappropriate encoding nucleic acid molecule under conditions suitable forexpression of the PRO5990 polypeptide and recovering the PRO5990polypeptide from the cell culture.

[0375] In yet another aspect, the invention concerns an isolated PRO5990polypeptide, comprising the sequence of amino acid residues from about 1or about 22 to about 468, inclusive, of FIG. 36 (SEQ ID NO:67), or afragment thereof which is biologically active or sufficient to provide abinding site for an anti-PRO5990 antibody, wherein the identification ofPRO5990 polypeptide fragments that possess biological activity orprovide a binding site for an anti-PRO5990 antibody may be accomplishedin a routine manner using techniques which are well known in the art.Preferably, the PRO5990 fragment retains a qualitative biologicalactivity of a native PRO5990 polypeptide.

[0376] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO5990 polypeptide havingthe sequence of amino acid residues from about 1 or about 22 to about468, inclusive, of FIG. 36 (SEQ ID NO:67), or (b) the complement of theDNA molecule of (a), and if the test DNA molecule has at least about an80% sequence identity, preferably at least about an 81% sequenceidentity, more preferably at least about an 82% sequence identity, yetmore preferably at least about an 83% sequence identity, yet morepreferably at least about an 84% sequence identity, yet more preferablyat least about an 85% sequence identity, yet more preferably at leastabout an 86% sequence identity, yet more preferably at least about an87% sequence identity, yet more preferably at least about an 88%sequence identity, yet more preferably at least about an 89% sequenceidentity, yet more preferably at least about a 90% sequence identity,yet more preferably at least about a 91% sequence identity, yet morepreferably at least about a 92% sequence identity, yet more preferablyat least about a 93% sequence identity, yet more preferably at leastabout a 94% sequence identity, yet more preferably at least about a 95%sequence identity, yet more preferably at least about a 96% sequenceidentity, yet more preferably at least about a 97% sequence identity,yet more preferably at least about a 98% sequence identity and yet morepreferably at least about a 99% sequence identity to (a) or (b), (ii)culturing a host cell comprising the test DNA molecule under conditionssuitable for expression of the polypeptide, and (iii) recovering thepolypeptide from the cell culture.

[0377] Another embodiment of the present invention is directed to theuse of a PRO5990 polypeptide, or an agonist or antagonist thereof asherein described, or an anti-PRO5990 antibody, for the preparation of amedicament useful in the treatment of a condition which is responsive tothe PRO5990 polypeptide, an agonist or antagonist thereof or ananti-PRO5990 antibody.

[0378] 19. PRO6030

[0379] A cDNA clone (designated herein as DNA96850-2705) has beenidentified that encodes a novel polypeptide, designated in the presentapplication as “PRO6030”.

[0380] In one embodiment, the invention provides an isolated nucleicacid molecule comprising a nucleotide sequence that encodes a PRO6030polypeptide.

[0381] In one aspect, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculeencoding a PRO6030 polypeptide having the sequence of amino acidresidues from about 1 or about 27 to about 322, inclusive, of FIG. 38(SEQ ID NO:72), or (b) the complement of the DNA molecule of (a).

[0382] In another aspect, the isolated nucleic acid molecule comprises(a) a nucleotide sequence encoding a PRO6030 polypeptide having thesequence of amino acid residues from about 1 or about 27 to about 322,inclusive, of FIG. 38 (SEQ ID NO:72), or (b) the complement of thenucleotide sequence of (a).

[0383] In other aspects, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculehaving the sequence of nucleotides from about 60 or about 138 to about1025, inclusive, of FIG. 37 (SEQ ID NO:71), or (b) the complement of theDNA molecule of (a).

[0384] In another aspect, the isolated nucleic acid molecule comprises(a) the nucleotide sequence of from about 60 or about 138 to about 1025,inclusive, of FIG. 37 (SEQ ID NO:71), or (b) the complement of thenucleotide sequence of (a).

[0385] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to (a) a DNAmolecule that encodes the same mature polypeptide encoded by the humanprotein cDNA deposited with the ATCC on Aug. 3, 1999 under ATCC DepositNo. 479-PTA (DNA96850-2705) or (b) the complement of the DNA molecule of(a). In a preferred embodiment, the isolated nucleic acid moleculecomprises (a) a nucleotide sequence encoding the same mature polypeptideencoded by the human protein cDNA deposited with the ATCC on Aug. 3,1999 under ATCC Deposit No. 479-PTA (DNA96850-2705) or (b) thecomplement of the nucleotide sequence of (a).

[0386] In another aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to (a) thefull-length polypeptide coding sequence of the human protein cDNAdeposited with the ATCC on Aug. 3, 1999 under ATCC Deposit No. 479-PTA(DNA96850-2705) or (b) the complement of the nucleotide sequence of (a).In a preferred embodiment, the isolated nucleic acid molecule comprises(a) the full-length polypeptide coding sequence of the DNA depositedwith the ATCC on Aug. 3, 1999 under ATCC Deposit No. 479-PTA(DNA96850-2705) or (b) the complement of the nucleotide sequence of (a).

[0387] In another aspect, the invention concerns an isolated nucleicacid molecule which encodes an active PRO6030 polypeptide as definedbelow comprising a nucleotide sequence that hybridizes to the complementof a nucleic acid sequence that encodes amino acids 1 or about 27 toabout 322, inclusive, of FIG. 38 (SEQ ID NO:72). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0388] In yet another aspect, the invention concerns an isolated nucleicacid molecule which encodes an active PRO6030 polypeptide as definedbelow comprising a nucleotide sequence that hybridizes to the complementof the nucleic acid sequence between about nucleotides 60 or about 138and about 1025, inclusive, of FIG. 37 (SEQ ID NO:71). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0389] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 528 nucleotides and which isproduced by hybridizing a test DNA molecule under stringent conditionswith (a) a DNA molecule encoding a PRO6030 polypeptide having thesequence of amino acid residues from about 1 or about 27 to about 322,inclusive, of FIG. 38 (SEQ ID NO:72), or (b) the complement of the DNAmolecule of (a), and, if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 81% sequence identity,more preferably at least about an 82% sequence identity, yet morepreferably at least about an 83% sequence identity, yet more preferablyat least about an 84% sequence identity, yet more preferably at leastabout an 85% sequence identity, yet more preferably at least about an86% sequence identity, yet more preferably at least about an 87%sequence identity, yet more preferably at least about an 88% sequenceidentity, yet more preferably at least about an 89% sequence identity,yet more preferably at least about a 90% sequence identity, yet morepreferably at least about a 91% sequence identity, yet more preferablyat least about a 92% sequence identity, yet more preferably at leastabout a 93% sequence identity, yet more preferably at least about a 94%sequence identity, yet more preferably at least about a 95% sequenceidentity, yet more preferably at least about a 96% sequence identity,yet more preferably at least about a 97% sequence identity, yet morepreferably at least about a 98% sequence identity and yet morepreferably at least about a 99% sequence identity to (a) or (b), andisolating the test DNA molecule.

[0390] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) a nucleotide sequence encoding apolypeptide scoring at least about 80% positives, preferably at leastabout 81% positives, more preferably at least about 82% positives, yetmore preferably at least about 83% positives, yet more preferably atleast about 84% positives, yet more preferably at least about 85%positives, yet more preferably at least about 86% positives, yet morepreferably at least about 87% positives, yet more preferably at leastabout 88% positives, yet more preferably at least about 89% positives,yet more preferably at least about 90% positives, yet more preferably atleast about 91% positives, yet more preferably at least about 92%positives, yet more preferably it least about 93% positives, yet morepreferably at least about 94% positives, yet more preferably at leastabout 95% positives, yet more preferably at least about 96% positives,yet more preferably at least about 97% positives, yet more preferably atleast about 98% positives and yet more preferably at least about 99%positives when compared with the amino acid sequence of residues about 1or about 27 to 322, inclusive, of FIG. 38 (SEQ ID NO:72), or (b) thecomplement of the nucleotide sequence of (a).

[0391] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO6030 polypeptide without theN-terminal signal sequence and/or the initiating methionine, or iscomplementary to such encoding nucleic acid molecule. The signal peptidehas been tentatively identified as extending from about amino acidposition 1 to about amino acid position 26 in the sequence of FIG. 38(SEQ ID NO:72). It is noted, however, that the C-terminal boundary ofthe signal peptide may vary, but most likely by no more than about 5amino acids on either side of the signal peptide C-terminal boundary asinitially identified herein, wherein the C-terminal boundary of thesignal peptide may be identified pursuant to criteria routinely employedin the art for identifying that type of amino acid sequence element(e.g., Nielsen et al., Prot. Eng. 10:1-6 (1997) and von Heinje et al.,Nucl. Acids. Res. 14:46834690 (1986)). Moreover, it is also recognizedthat, in some cases, cleavage of a signal sequence from a secretedpolypeptide is not entirely uniform, resulting in more than one secretedspecies. These polypeptides, and the polynucleotides encoding them, arecontemplated by the present invention. As such, for purposes of thepresent application, the signal peptide of the PRO6030 polypeptide shownin FIG. 38 (SEQ ID NO:72) extends from amino acids 1 to X of FIG. 38(SEQ ID NO:72), wherein X is any amino acid from 21 to 31 of FIG. 38(SEQ ID NO:72). Therefore, mature forms of the PRO6030 polypeptide whichare encompassed by the present invention include those comprising aminoacids X to 322 of FIG. 38 (SEQ ID NO:72), wherein X is any amino acidfrom 21 to 31 of FIG. 38 (SEQ ID NO:72) and variants thereof asdescribed below. Isolated nucleic acid molecules encoding thesepolypeptides are also contemplated.

[0392] Another aspect the invention provides an isolated nucleic acidmolecule comprising a nucleotide sequence encoding a PRO6030 polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated, or is complementary to such encoding nucleotidesequence, wherein the transmembrane domain has been tentativelyidentified as extending from about amino acid position 142 to aboutamino acid position 158 in the sequence of FIG. 38 (SEQ ID NO:72).Therefore, soluble extracellular domains of the herein described PRO6030polypeptides are contemplated.

[0393] In this regard, another aspect of the present invention isdirected to an isolated nucleic acid molecule which comprises anucleotide sequence having at least about 80% sequence identity,preferably at least about 81% sequence identity, more preferably atleast about 82% sequence identity, yet more preferably at least about83% sequence identity, yet more preferably at least about 84% sequenceidentity, yet more preferably at least about 85% sequence identity, yetmore preferably at least about 86% sequence identity, yet morepreferably at least about 87% sequence identity, yet more preferably atleast about 88% sequence identity, yet more preferably at least about89% sequence identity, yet more preferably at least about 90% sequenceidentity, yet more preferably at least about 91% sequence identity, yetmore preferably at least about 92% sequence identity, yet morepreferably at least about 93% sequence identity, yet more preferably atleast about 94% sequence identity, yet more preferably at least about95% sequence identity, yet more preferably at least about 96% sequenceidentity, yet more preferably at least about 97% sequence identity, yetmore preferably at least about 98% sequence identity and yet morepreferably at least about 99% sequence identity to (a) a DNA moleculeencoding amino acids 1 to X of FIG. 38 (SEQ ID NO:72), where X is anyamino acid from 137 to 147 of FIG. 38 (SEQ ID NO:72), or (b) thecomplement of the DNA molecule of (a). In a specific aspect, theisolated nucleic acid molecule comprises a nucleotide sequence which (a)encodes amino acids 1 to X of FIG. 38 (SEQ ID NO:72), where X is anyamino acid from 137 to 147 of FIG. 38 (SEQ ID NO:72), or (b) is thecomplement of the DNA molecule of (a).

[0394] In yet another aspect of the present invention, the isolatednucleic acid molecule (a) encodes a polypeptide scoring at least about80% positives, preferably at least about 81% positives, more preferablyat least about 82% positives, yet more preferably at least about 83%positives, yet more preferably at least about 84% positives, yet morepreferably at least about 85% positives, yet more preferably at leastabout 86% positives, yet more preferably at least about 87% positives,yet more preferably at least about 88% positives, yet more preferably atleast about 89% positives, yet more preferably at least about 90%positives, yet more preferably at least about 91% positives, yet morepreferably at least about 92% positives, yet more preferably at leastabout 93% positives, yet more preferably at least about 94% positives,yet more preferably at least about 95% positives, yet more preferably atleast about 96% positives, yet more preferably at least about 97%positives, yet more preferably at least about 98% positives and yet morepreferably at least about 99% positives when compared with the aminoacid sequence of residues about 1 to X of FIG. 38 (SEQ ID NO:72), whereX is any amino acid from 137 to 147 of FIG. 38 (SEQ ID NO:72), or (b) isthe complement of the DNA molecule of (a).

[0395] Another embodiment is directed to fragments of a PRO6030polypeptide coding sequence that may find use as, for example,hybridization probes or for encoding fragments of a PRO6030 polypeptidethat may optionally encode a polypeptide comprising a binding site foran anti-PRO6030 antibody. Such nucleic acid fragments are usually atleast about 20 nucleotides in length, preferably at least about 30nucleotides in length, more preferably at least about 40 nucleotides inlength, yet more preferably at least about 50 nucleotides in length, yetmore preferably at least about 60 nucleotides in length, yet morepreferably at least about 70 nucleotides in length, yet more preferablyat least about 80 nucleotides in length, yet more preferably at leastabout 90 nucleotides in length, yet more preferably at least about 100nucleotides in length, yet more preferably at least about 110nucleotides in length, yet more preferably at least about 120nucleotides in length, yet more preferably at least about 130nucleotides in length, yet more preferably at least about 140nucleotides in length, yet more preferably at least about 150nucleotides in length, yet more preferably at least about 160nucleotides in length, yet more preferably at least about 170nucleotides in length, yet more preferably at least about 180nucleotides in length, yet more preferably at least about 190nucleotides in length, yet more preferably at least about 200nucleotides in length, yet more preferably at least about 250nucleotides in length, yet more preferably at least about 300nucleotides in length, yet more preferably at least about 350nucleotides in length, yet more preferably at least about 400nucleotides in length, yet more preferably at least about 450nucleotides in length, yet more preferably at least about 500nucleotides in length, yet more preferably at least about 600nucleotides in length, yet more preferably at least about 700nucleotides in length, yet more preferably at least about 800nucleotides in length, yet more preferably at least about 900nucleotides in length and yet more preferably at least about 1000nucleotides in length, wherein in this context the term “about” meansthe referenced nucleotide sequence length plus or minus 10% of thatreferenced length. In a preferred embodiment, the nucleotide sequencefragment is derived from any coding region of the nucleotide sequenceshown in FIG. 37 (SEQ ID NO:71). It is noted that novel fragments of aPRO6030 polypeptide-encoding nucleotide sequence may be determined in aroutine manner by aligning the PRO6030 polypeptide-encoding nucleotidesequence with other known nucleotide sequences using any of a number ofwell known sequence alignment programs and determining which PRO6030polypeptide-encoding nucleotide sequence fragment(s) are novel. All ofsuch PRO6030 polypeptide-encoding nucleotide sequences are contemplatedherein and can be determined without undue experimentation. Alsocontemplated are the PRO6030 polypeptide fragments encoded by thesenucleotide molecule fragments, preferably those PRO6030 polypeptidefragments that comprise a binding site for an anti-PRO6030 antibody.

[0396] In another embodiment, the invention provides isolated PRO6030polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0397] In a specific aspect, the invention provides isolated nativesequence PRO6030 polypeptide, which in certain embodiments, includes anamino acid sequence comprising residues from about 1 or about 27 toabout 322 of FIG. 38 (SEQ ID NO:72).

[0398] In another aspect, the invention concerns an isolated PRO6030polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to the sequenceof amino acid residues from about 1 or about 27 to about 322, inclusive,of FIG. 38 (SEQ ID NO:72).

[0399] In a further aspect, the invention concerns an isolated PRO6030polypeptide comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 81% sequence identity, morepreferably at least about 82% sequence identity, yet more preferably atleast about 83% sequence identity, yet more preferably at least about84% sequence identity, yet more preferably at least about 85% sequenceidentity, yet more preferably at least about 86% sequence identity, yetmore preferably at least about 87% sequence identity, yet morepreferably at least about 88% sequence identity, yet more preferably atleast about 89% sequence identity, yet more preferably at least about90% sequence identity, yet more preferably at least about 91% sequenceidentity, yet more preferably at least about 92% sequence identity, yetmore preferably at least about 93% sequence identity, yet morepreferably at least about 94% sequence identity, yet more preferably atleast about 95% sequence identity, yet more preferably at least about96% sequence identity, yet more preferably at least about 97% sequenceidentity, yet more preferably at least about 98% sequence identity andyet more preferably at least about 99% sequence identity to an aminoacid sequence encoded by the human protein cDNA deposited with the ATCCon Aug. 3, 1999 under ATCC Deposit No. 479-PTA (DNA96850-2705). In apreferred embodiment, the isolated PRO6030 polypeptide comprises anamino acid sequence encoded by the human protein cDNA deposited with theATCC on Aug. 3, 1999 under ATCC Deposit No. 479-PTA (DNA96850-2705).

[0400] In a further aspect, the invention concerns an isolated PRO6030polypeptide comprising an amino acid sequence scoring at least about 80%positives, preferably at least about 81% positives, more preferably atleast about 82% positives, yet more preferably at least about 83%positives, yet more preferably at least about 84% positives, yet morepreferably at least about 85% positives, yet more preferably at leastabout 86% positives, yet more preferably at least about 87% positives,yet more preferably at least about 88% positives, yet more preferably atleast about 89% positives, yet more preferably at least about 90%positives, yet more preferably at least about 91% positives, yet morepreferably at least about 92% positives, yet more preferably at leastabout 93% positives, yet more preferably at least about 94% positives,yet more preferably at least about 95% positives, yet more preferably atleast about 96% positives, yet more preferably at least about 97%positives, yet more preferably at least about 98% positives and yet morepreferably at least about 99% positives when compared with the aminoacid sequence of residues from about 1 or about 27 to about 322,inclusive, of FIG. 38 (SEQ ID NO:72).

[0401] In a specific aspect, the invention provides an isolated PRO6030polypeptide without the N-terminal signal sequence and/or the initiatingmethionine and is encoded by a nucleotide sequence that encodes such anamino acid sequence as hereinbefore described. Processes for producingthe same are also herein described, wherein those processes compriseculturing a host cell comprising a vector which comprises theappropriate encoding nucleic acid molecule under conditions suitable forexpression of the PRO6030 polypeptide and recovering the PRO6030polypeptide from the cell culture.

[0402] Another aspect the invention provides an isolated PRO6030polypeptide which is either transmembrane domain-deleted ortransmembrane domain-inactivated. Processes for producing the same arealso herein described, wherein those processes comprise culturing a hostcell comprising a vector which comprises the appropriate encodingnucleic acid molecule under conditions suitable for expression of thePRO6030 polypeptide and recovering the PRO6030 polypeptide from the cellculture.

[0403] As such, one aspect of the present invention is directed to anisolated soluble PRO6030 polypeptide which comprises an amino acidsequence having at least about 80% sequence identity, preferably atleast about 81% sequence identity, more preferably at least about 82%sequence identity, yet more preferably at least about 83% sequenceidentity, yet more preferably at least about 84% sequence identity, yetmore preferably at least about 85% sequence identity, yet morepreferably at least about 86% sequence identity, yet more preferably atleast about 87% sequence identity, yet more preferably at least about88% sequence identity, yet more preferably at least about 89% sequenceidentity, yet more preferably at least about 90% sequence identity, yetmore preferably at least about 91% sequence identity, yet morepreferably at least about 92% sequence identity, yet more preferably atleast about 93% sequence identity, yet more preferably at least about94% sequence identity, yet more preferably at least about 95% sequenceidentity, yet more preferably at least about 96% sequence identity, yetmore preferably at least about 97% sequence identity, yet morepreferably at least about 98% sequence identity and yet more preferablyat least about 99% sequence identity to amino acids 1 to X of FIG. 38(SEQ ID NO:72), where X is any amino acid from 137 to 147 of FIG. 38(SEQ ID NO:72). In a preferred aspect, the isolated soluble PRO6030polypeptide comprises amino acids 1 to X of FIG. 38 (SEQ ID NO:72),where X is any amino acid from 137 to 147 of FIG. 38 (SEQ ID NO:72).

[0404] In yet another aspect of the present invention, the isolatedsoluble PRO6030 polypeptide comprises an amino acid sequence whichscores at least about 80% positives, preferably at least about 81%positives, more preferably at least about 82% positives, yet morepreferably at least about 83% positives, yet more preferably at leastabout 84% positives, yet more preferably at least about 85% positives,yet more preferably at least about 86% positives, yet more preferably atleast about 87% positives, yet more preferably at least about 88%positives, yet more preferably at least about 89% positives, yet morepreferably at least about 90% positives, yet more preferably at leastabout 91% positives, yet more preferably at least about 92% positives,yet more preferably at least about 93% positives, yet more preferably atleast about 94% positives, yet more preferably at least about 95%positives, yet more preferably at least about 96% positives, yet morepreferably at least about 97% positives, yet more preferably at leastabout 98% positives and yet more preferably at least about 99% positiveswhen compared with the amino acid sequence of residues about 1 to X ofFIG. 38 (SEQ ID NO:72), where X is any amino acid from 137 to 147 ofFIG. 38 (SEQ ID NO:72).

[0405] In yet another aspect, the invention concerns an isolated PRO6030polypeptide, comprising the sequence of amino acid residues from about 1or about 27 to about 322, inclusive, of FIG. 38 (SEQ ID NO:72), or afragment thereof which is biologically active or sufficient to provide abinding site for an anti-PRO6030 antibody, wherein the identification ofPRO6030 polypeptide fragments that possess biological activity orprovide a binding site for an anti-PRO6030 antibody may be accomplishedin a routine manner using techniques which are well known in the art.Preferably, the PRO6030 fragment retains a qualitative biologicalactivity of a native PRO6030 polypeptide.

[0406] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO6030 polypeptide havingthe sequence of amino acid residues from about 1 or about 27 to about322, inclusive, of FIG. 38 (SEQ ID NO:72), or (b) the complement of theDNA molecule of (a), and if the test DNA molecule has at least about an80% sequence identity, preferably at least about an 81% sequenceidentity, more preferably at least about an 82% sequence identity, yetmore preferably at least about an 83% sequence identity, yet morepreferably at least about an 84% sequence identity, yet more preferablyat least about an 85% sequence identity, yet more preferably at leastabout an 86% sequence identity, yet more preferably at least about an87% sequence identity, yet more preferably at least about an 88%sequence identity, yet more preferably at least about an 89% sequenceidentity, yet more preferably at least about a 90% sequence identity,yet more preferably at least about a 91% sequence identity, yet morepreferably at least about a 92% sequence identity, yet more preferablyat least about a 93% sequence identity, yet more preferably at leastabout a 94% sequence identity, yet more preferably at least about a 95%sequence identity, yet more preferably at least about a 96% sequenceidentity, yet more preferably at least about a 97% sequence identity,yet more preferably at least about a 98% sequence identity and yet morepreferably at least about a 99% sequence identity to (a) or (b), (ii)culturing a host cell comprising the test DNA molecule under conditionssuitable for expression of the polypeptide, and (iii) recovering thepolypeptide from the cell culture.

[0407] In a still further embodiment, the invention concerns acomposition of matter comprising a PRO6030 polypeptide, or an agonist orantagonist of a PRO6030 polypeptide as herein described, or ananti-PRO6030 antibody, in combination with a carrier. Optionally, thecarrier is a pharmaceutically acceptable carrier.

[0408] Another embodiment of the present invention is directed to theuse of a PRO6030 polypeptide, or an agonist or antagonist thereof asherein described, or an anti-PRO6030 antibody, for the preparation of amedicament useful in the treatment of a condition which is responsive tothe PRO6030 polypeptide, an agonist or antagonist thereof or ananti-PRO6030 antibody.

[0409] 20. PRO4424

[0410] A cDNA clone (DNA96857-2636) has been identified that encodes anovel polypeptide having homology to a protein in GenBank, accessionnumber HGS_A135 and designated in the present application as “PRO4424”.

[0411] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4424 polypeptide.

[0412] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4424 polypeptide having the sequence of aminoacid residues from 1 or about 29 to about 221, inclusive of FIG. 40 (SEQID NO:74), or (b) the complement of the DNA molecule of (a).

[0413] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4424 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 136 andabout 714, inclusive, of FIG. 39 (SEQ ID NO:73). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0414] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 17-PTA (DNA96857-2636), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 17-PTA (DNA96857-2636).

[0415] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 29 to about 221, inclusive of FIG. 40(SEQ ID NO:74), or the complement of the DNA of (a).

[0416] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4424 polypeptide having the sequence of amino acid residues fromabout 29 to about 221, inclusive of FIG. 40 (SEQ ID NO:74), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0417] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 29to about 221, inclusive of FIG. 40 (SEQ ID NO:74), or (b) the complementof the DNA of (a).

[0418] Another embodiment is directed to fragments of a PRO4424polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0419] In another embodiment, the invention provides isolated PRO4424polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0420] In a specific aspect, the invention provides isolated nativesequence PRO4424 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 29 through 221 of FIG. 40 (SEQ IDNO:74).

[0421] In another aspect, the invention concerns an isolated PRO4424polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues29 to about 221, inclusive of FIG. 40 (SEQ ID NO:74).

[0422] In a further aspect, the invention concerns an isolated PRO4424polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 29through 221 of FIG. 40 (SEQ ID NO:74).

[0423] In yet another aspect, the invention concerns an isolated PRO4424polypeptide, comprising the sequence of amino acid residues 29 to about221, inclusive of FIG. 40 (SEQ ID NO:74), or a fragment thereofsufficient to provide a binding site for an anti-PRO4424 antibody.Preferably, the PRO4424 fragment retains a qualitative biologicalactivity of a native PRO4424 polypeptide.

[0424] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4424 polypeptide havingthe sequence of amino acid residues from about 29 to about 221,inclusive of FIG. 40 (SEQ ID NO:74), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0425] 21. PRO4422

[0426] A cDNA clone (DNA96867-2620) has been identified that encodes anovel polypeptide having homology to lysozyme g and designated in thepresent application as “PRO4422”.

[0427] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4422 polypeptide.

[0428] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4422 polypeptide having the sequence of aminoacid residues from 1 or about 20 to about 194, inclusive of FIG. 42 (SEQID NO:76), or (b) the complement of the DNA molecule of (a).

[0429] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4422 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 375 andabout 899, inclusive, of FIG. 41 (SEQ ID NO:75). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0430] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 203972 (DNA96867-2620), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 203972 (DNA96867-2620).

[0431] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 20 to about 194, inclusive of FIG. 42(SEQ ID NO:76), or the complement of the DNA of (a).

[0432] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4422 polypeptide having the sequence of amino acid residues fromabout 20 to about 194, inclusive of FIG. 42 (SEQ ID NO:76), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0433] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 20to about 194, inclusive of FIG. 42 (SEQ ID NO:76), or (b) the complementof the DNA of (a).

[0434] Another embodiment is directed to fragments of a PRO4422polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0435] In another embodiment, the invention provides isolated PRO4422polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0436] In a specific aspect, the invention provides isolated nativesequence PRO4422 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 20 through 194 of FIG. 42 (SEQ IDNO:76).

[0437] In another aspect, the invention concerns an isolated PRO4422polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues20 to about 194, inclusive of FIG. 42 (SEQ ID NO:76).

[0438] In a further aspect, the invention concerns an isolated PRO4422polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 20through 194 of FIG. 42 (SEQ ID NO:76).

[0439] In yet another aspect, the invention concerns an isolated PRO4422polypeptide, comprising the sequence of amino acid residues 20 to about194, inclusive of FIG. 42 (SEQ ID NO:76), or a fragment thereofsufficient to provide a binding site for an anti-PRO4422 antibody.Preferably, the PRO4422 fragment retains a qualitative biologicalactivity of a native PRO4422 polypeptide.

[0440] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4422 polypeptide havingthe sequence of amino acid residues from about 20 to about 194,inclusive of FIG. 42 (SEQ ID NO:76), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0441] 22. PRO4430

[0442] A cDNA clone (DNA96878-2626) has been identified that encodes anovel polypeptide having homology to a protein in GenBank, accessionnumber MMHC213L3_(—)9, and designated in the present application as“PRO4430”.

[0443] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4430 polypeptide.

[0444] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4430 polypeptide having the sequence of aminoacid residues from 1 or about 19 to about 125, inclusive of FIG. 44 (SEQID NO:78), or (b) the complement of the DNA molecule of (a).

[0445] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4430 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 110 andabout 430, inclusive, of FIG. 43 (SEQ ID NO:77). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0446] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 23-PTA (DNA96878-2626), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 23-PTA (DNA96878-2626).

[0447] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 19 to about 125, inclusive of FIG. 44(SEQ ID NO:78), or the complement of the DNA of (a).

[0448] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4430 polypeptide having the sequence of amino acid residues fromabout 19 to about 125, inclusive of FIG. 44 (SEQ ID NO:78), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0449] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 19to about 125, inclusive of FIG. 44 (SEQ ID NO:78), or (b) the complementof the DNA of (a).

[0450] Another embodiment is directed to fragments of a PRO4430polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0451] In another embodiment, the invention provides isolated PRO4430polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0452] In a specific aspect, the invention provides isolated nativesequence PRO4430 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 19 through 125 of FIG. 44 (SEQ IDNO:78).

[0453] In another aspect, the invention concerns an isolated PRO4430polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues19 to about 125, inclusive of FIG. 44 (SEQ ID NO:78).

[0454] In a further aspect, the invention concerns an isolated PRO4430polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 19through 125 of FIG. 44 (SEQ ID NO:78).

[0455] In yet another aspect, the invention concerns an isolated PRO4430polypeptide, comprising the sequence of amino acid residues 19 to about125, inclusive of FIG. 44 (SEQ ID NO:78), or a fragment thereofsufficient to provide a binding site for an anti-PRO4430 antibody.Preferably, the PRO4430 fragment retains a qualitative biologicalactivity of a native PRO4430 polypeptide.

[0456] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4430 polypeptide havingthe sequence of amino acid residues from about 19 to about 125,inclusive of FIG. 44 (SEQ ID NO:78), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture. 23. PRO4499

[0457] A cDNA clone (DNA96889-2641) has been identified that encodes anovel polypeptide and designated in the present application as“PRO4499”.

[0458] In one embodiment, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4499 polypeptide.

[0459] In one aspect, the isolated nucleic acid comprises DNA having atleast about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to (a) a DNAmolecule encoding a PRO4499 polypeptide having the sequence of aminoacid residues from 1 or about 31 to about 339, inclusive of FIG. 46 (SEQID NO:80), or (b) the complement of the DNA molecule of (a).

[0460] In another aspect, the invention concerns an isolated nucleicacid molecule encoding a PRO4499 polypeptide comprising DNA hybridizingto the complement of the nucleic acid between about residues 275 andabout 1201, inclusive, of FIG. 45 (SEQ ID NO:79). Preferably,hybridization occurs under stringent hybridization and wash conditions.

[0461] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising DNA having at least about 80% sequenceidentity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to (a) a DNA molecule encoding thesame mature polypeptide encoded by the human protein cDNA in ATCCDeposit No. 119-PTA (DNA96889-2641), or (b) the complement of the DNAmolecule of (a). In a preferred embodiment, the nucleic acid comprises aDNA encoding the same mature polypeptide encoded by the human proteincDNA in ATCC Deposit No. 119-PTA (DNA96889-2641).

[0462] In a still further aspect, the invention concerns an isolatednucleic acid molecule comprising (a) DNA encoding a polypeptide havingat least about 80% sequence identity, preferably at least about 85%sequence identity, more preferably at least about 90% sequence identity,most preferably at least about 95% sequence identity to the sequence ofamino acid residues from about 31 to about 339, inclusive of FIG. 46(SEQ ID NO:80), or the complement of the DNA of (a).

[0463] In a further aspect, the invention concerns an isolated nucleicacid molecule having at least about 50 nucleotides, and preferably atleast about 100 nucleotides and produced by hybridizing a test DNAmolecule under stringent conditions with (a) a DNA molecule encoding aPRO4499 polypeptide having the sequence of amino acid residues fromabout 31 to about 339, inclusive of FIG. 46 (SEQ ID NO:80), or (b) thecomplement of the DNA molecule of (a), and, if the DNA molecule has atleast about an 80% sequence identity, preferably at least about an 85%sequence identity, more preferably at least about a 90% sequenceidentity, most preferably at least about a 95% sequence identity to (a)or (b), isolating the test DNA molecule.

[0464] In a specific aspect, the invention provides an isolated nucleicacid molecule comprising DNA encoding a PRO4499 polypeptide, with orwithout the N-terminal signal sequence and/or the initiating methionine,and its soluble variants (i.e. transmembrane domain deleted orinactivated), or is complementary to such encoding nucleic acidmolecule. The signal peptide has been tentatively identified asextending from amino acid position 1 through about amino acid position30 in the sequence of FIG. 46 (SEQ ID NO:80). The transmembrane domainhas been tentatively identified as extending from about amino acidposition 171 through about amino acid position 190 in the PRO4499 aminoacid sequence (FIG. 46, SEQ ID NO:80).

[0465] In another aspect, the invention concerns an isolated nucleicacid molecule comprising (a) DNA encoding a polypeptide scoring at leastabout 80% positives, preferably at least about 85% positives, morepreferably at least about 90% positives, most preferably at least about95% positives when compared with the amino acid sequence of residues 31to about 339, inclusive of FIG. 46 (SEQ ID NO:80), or (b) the complementof the DNA of (a).

[0466] Another embodiment is directed to fragments of a PRO4499polypeptide coding sequence that may find use as hybridization probes.Such nucleic acid fragments are from about 20 through about 80nucleotides in length, preferably from about 20 through about 60nucleotides in length, more preferably from about 20 through about 50nucleotides in length, and most preferably from about 20 through about40 nucleotides in length.

[0467] In another embodiment, the invention provides isolated PRO4499polypeptide encoded by any of the isolated nucleic acid sequenceshereinabove defined.

[0468] In a specific aspect, the invention provides isolated nativesequence PRO4499 polypeptide, which in one embodiment, includes an aminoacid sequence comprising residues 31 through 339 of FIG. 46 (SEQ IDNO:80).

[0469] In another aspect, the invention concerns an isolated PRO4499polypeptide, comprising an amino acid sequence having at least about 80%sequence identity, preferably at least about 85% sequence identity, morepreferably at least about 90% sequence identity, most preferably atleast about 95% sequence identity to the sequence of amino acid residues31 to about 339, inclusive of FIG. 46 (SEQ ID NO:80).

[0470] In a further aspect, the invention concerns an isolated PRO4499polypeptide, comprising an amino acid sequence scoring at least about80% positives, preferably at least about 85% positives, more preferablyat least about 90% positives, most preferably at least about 95%positives when compared with the amino acid sequence of residues 31through 339 of FIG. 46 (SEQ ID NO:80).

[0471] In yet another aspect, the invention concerns an isolated PRO4499polypeptide, comprising the sequence of amino acid residues 31 to about339, inclusive of FIG. 46 (SEQ ID NO:80), or a fragment thereofsufficient to provide a binding site for an anti-PRO4499 antibody.Preferably, the PRO4499 fragment retains a qualitative biologicalactivity of a native PRO4499 polypeptide.

[0472] In a still further aspect, the invention provides a polypeptideproduced by (i) hybridizing a test DNA molecule under stringentconditions with (a) a DNA molecule encoding a PRO4499 polypeptide havingthe sequence of amino acid residues from about 31 to about 339,inclusive of FIG. 46 (SEQ ID NO:80), or (b) the complement of the DNAmolecule of (a), and if the test DNA molecule has at least about an 80%sequence identity, preferably at least about an 85% sequence identity,more preferably at least about a 90% sequence identity, most preferablyat least about a 95% sequence identity to (a) or (b), (ii) culturing ahost cell comprising the test DNA molecule under conditions suitable forexpression of the polypeptide, and (iii) recovering the polypeptide fromthe cell culture.

[0473] 24. Additional Embodiments

[0474] In other embodiments of the present invention, the inventionprovides vectors comprising DNA encoding any of the herein describedpolypeptides. Host cell comprising any such vector are also provided. Byway of example, the host cells may be CHO cells, E. coli, or yeast. Aprocess for producing any of the herein described polypeptides isfurther provided and comprises culturing host cells under conditionssuitable for expression of the desired polypeptide and recovering thedesired polypeptide from the cell culture.

[0475] In other embodiments, the invention provides chimeric moleculescomprising any of the herein described polypeptides fused to aheterologous polypeptide or amino acid sequence. Example of suchchimeric molecules comprise any of the herein described polypeptidesfused to an epitope tag sequence or a Fc region of an immunoglobulin.

[0476] In another embodiment, the invention provides an antibody whichspecifically binds to any of the above or below described polypeptides.Optionally, the antibody is a monoclonal antibody, humanized antibody,antibody fragment or single-chain antibody.

[0477] In yet other embodiments, the invention provides oligonucleotideprobes useful for isolating genomic and cDNA nucleotide sequences or asantisense probes, wherein those probes may be derived from any of theabove or below described nucleotide sequences.

[0478] In other embodiments, the invention provides an isolated nucleicacid molecule comprising a nucleotide sequence that encodes a PROpolypeptide.

[0479] In one aspect, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% nucleic acid sequenceidentity, alternatively at least about 81% nucleic acid sequenceidentity, alternatively at least about 82% nucleic acid sequenceidentity, alternatively at least about 83% nucleic acid sequenceidentity, alternatively at least about 84% nucleic acid sequenceidentity, alternatively at least about 85% nucleic acid sequenceidentity, alternatively at least about 86% nucleic acid sequenceidentity, alternatively at least about 87% nucleic acid sequenceidentity, alternatively at least about 88% nucleic acid sequenceidentity, alternatively at least about 89% nucleic acid sequenceidentity, alternatively at least about 90% nucleic acid sequenceidentity, alternatively at least about 91% nucleic acid sequenceidentity, alternatively at least about 92% nucleic acid sequenceidentity, alternatively at least about 93% nucleic acid sequenceidentity, alternatively at least about 94% nucleic acid sequenceidentity, alternatively at least about 95% nucleic acid sequenceidentity, alternatively at least about 96% nucleic acid sequenceidentity, alternatively at least about 97% nucleic acid sequenceidentity, alternatively at least about 98% nucleic acid sequenceidentity and alternatively at least about 99% nucleic acid sequenceidentity to (a) a DNA molecule encoding a PRO polypeptide having afull-length amino acid sequence as disclosed herein, an amino acidsequence lacking the signal peptide as disclosed herein, anextracellular domain of a transmembrane protein, with or without thesignal peptide, as disclosed herein or any other specifically definedfragment of the full-length amino acid sequence as disclosed herein, or(b) the complement of the DNA molecule of (a).

[0480] In other aspects, the isolated nucleic acid molecule comprises anucleotide sequence having at least about 80% nucleic acid sequenceidentity, alternatively at least about 81% nucleic acid sequenceidentity, alternatively at least about 82% nucleic acid sequenceidentity, alternatively at least about 83% nucleic acid sequenceidentity, alternatively at least about 84% nucleic acid sequenceidentity, alternatively at least about 85% nucleic acid sequenceidentity, alternatively at least about 86% nucleic acid sequenceidentity, alternatively at least about 87% nucleic acid sequenceidentity, alternatively at least about 88% nucleic acid sequenceidentity, alternatively at least about 89% nucleic acid sequenceidentity, alternatively at least about 90% nucleic acid sequenceidentity, alternatively at least about 91% nucleic acid sequenceidentity, alternatively at least about 92% nucleic acid sequenceidentity, alternatively at least about 93% nucleic acid sequenceidentity, alternatively at least about 94% nucleic acid sequenceidentity, alternatively at least about 95% nucleic acid sequenceidentity, alternatively at least about 96% nucleic acid sequenceidentity, alternatively at least about 97% nucleic acid sequenceidentity, alternatively at least about 98% nucleic acid sequenceidentity and alternatively at least about 99% nucleic acid sequenceidentity to (a) a DNA molecule comprising the coding sequence of afull-length PRO polypeptide cDNA as disclosed herein, the codingsequence of a PRO polypeptide lacking the signal peptide as disclosedherein, the coding sequence of an extracellular domain of atransmembrane PRO polypeptide, with or without the signal peptide, asdisclosed herein or the coding sequence of any other specificallydefined fragment of the full-length amino acid sequence as disclosedherein, or (b) the complement of the DNA molecule of (a).

[0481] In a further aspect, the invention concerns an isolated nucleicacid molecule comprising a nucleotide sequence having at least about 80%nucleic acid sequence identity, alternatively at least about 81% nucleicacid sequence identity, alternatively at least about 82% nucleic acidsequence identity, alternatively at least about 83% nucleic acidsequence identity, alternatively at least about 84% nucleic acidsequence identity, alternatively at least about 85% nucleic acidsequence identity, alternatively at least about 86% nucleic acidsequence identity, alternatively at least about 87% nucleic acidsequence identity, alternatively at least about 88% nucleic acidsequence identity, alternatively at least about 89% nucleic acidsequence identity, alternatively at least about 90% nucleic acidsequence identity, alternatively at least about 91% nucleic acidsequence identity, alternatively at least about 92% nucleic acidsequence identity, alternatively at least about 93% nucleic acidsequence identity, alternatively at least about 94% nucleic acidsequence identity, alternatively at least about 95% nucleic acidsequence identity, alternatively at least about 96% nucleic acidsequence identity, alternatively at least about 97% nucleic acidsequence identity, alternatively at least about 98% nucleic acidsequence identity and alternatively at least about 99% nucleic acidsequence identity to (a) a DNA molecule that encodes the same maturepolypeptide encoded by any of the human protein cDNAs deposited with theATCC as disclosed herein, or (b) the complement of the DNA molecule of(a).

[0482] Another aspect the invention provides an isolated nucleic acidmolecule comprising a nucleotide sequence encoding a PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated, or is complementary to such encoding nucleotidesequence, wherein the transmembrane domain(s) of such polypeptide aredisclosed herein. Therefore, soluble extracellular domains of the hereindescribed PRO polypeptides are contemplated.

[0483] Another embodiment is directed to fragments of a PRO polypeptidecoding sequence, or the complement thereof, that may find use as, forexample, hybridization probes, for encoding fragments of a PROpolypeptide that may optionally encode a polypeptide comprising abinding site for an anti-PRO antibody or as antisense oligonucleotideprobes. Such nucleic acid fragments are usually at least about 20nucleotides in length, alternatively at least about 30 nucleotides inlength, alternatively at least about 40 nucleotides in length,alternatively at least about 50 nucleotides in length, alternatively atleast about 60 nucleotides in length, alternatively at least about 70nucleotides in length, alternatively at least about 80 nucleotides inlength, alternatively at least about 90 nucleotides in length,alternatively at least about 100 nucleotides in length, alternatively atleast about 110 nucleotides in length, alternatively at least about 120nucleotides in length, alternatively at least about 130 nucleotides inlength, alternatively at least about 140 nucleotides in length,alternatively at least about 150 nucleotides in length, alternatively atleast about 160 nucleotides in length, alternatively at least about 170nucleotides in length, alternatively at least about 180 nucleotides inlength, alternatively at least about 190 nucleotides in length,alternatively at least about 200 nucleotides in length, alternatively atleast about 250 nucleotides in length, alternatively at least about 300nucleotides in length, alternatively at least about 350 nucleotides inlength, alternatively at least about 400 nucleotides in length,alternatively at least about 450 nucleotides in length, alternatively atleast about 500 nucleotides in length, alternatively at least about 600nucleotides in length, alternatively at least about 700 nucleotides inlength, alternatively at least about 800 nucleotides in length,alternatively at least about 900 nucleotides in length and alternativelyat least about 1000 nucleotides in length, wherein in this context theterm “about” means the referenced nucleotide sequence length plus orminus 10% of that referenced length. It is noted that novel fragments ofa PRO polypeptide-encoding nucleotide sequence may be determined in aroutine manner by aligning the PRO polypeptide-encoding nucleotidesequence with other known nucleotide sequences using any of a number ofwell known sequence alignment programs and determining which PROpolypeptide-encoding nucleotide sequence fragment(s) are novel. All ofsuch PRO polypeptide-encoding nucleotide sequences are contemplatedherein. Also contemplated are the PRO polypeptide fragments encoded bythese nucleotide molecule fragments, preferably those PRO polypeptidefragments that comprise a binding site for an anti-PRO antibody.

[0484] In another embodiment, the invention provides isolated PROpolypeptide encoded by any of the isolated nucleic acid sequenceshereinabove identified.

[0485] In a certain aspect, the invention concerns an isolated PROpolypeptide, comprising an amino acid sequence having at least about 80%amino acid sequence identity, alternatively at least about 81% aminoacid sequence identity, alternatively at least about 82% amino acidsequence identity, alternatively at least about 83% amino acid sequenceidentity, alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to a PROpolypeptide having a full-length amino acid sequence as disclosedherein, an amino acid sequence lacking the signal peptide as disclosedherein, an extracellular domain of a transmembrane protein, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of the full-length amino acid sequence asdisclosed herein.

[0486] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence having at least about 80%amino acid sequence identity, alternatively at least about 81% aminoacid sequence identity, alternatively at least about 82% amino acidsequence identity, alternatively at least about 83% amino acid sequenceidentity, alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to anamino acid sequence encoded by any of the human protein cDNAs depositedwith the ATCC as disclosed herein.

[0487] In a further aspect, the invention concerns an isolated PROpolypeptide comprising an amino acid sequence scoring at least about 80%positives, alternatively at least about 81% positives, alternatively atleast about 82% positives, alternatively at least about 83% positives,alternatively at least about 84% positives, alternatively at least about85% positives, alternatively at least about 86% positives, alternativelyat least about 87% positives, alternatively at least about 88%positives, alternatively at least about 89% positives, alternatively atleast about 90% positives, alternatively at least about 91% positives,alternatively at least about 92% positives, alternatively at least about93% positives, alternatively at least about 94% positives, alternativelyat least about 95% positives, alternatively at least about 96%positives, alternatively at least about 97% positives, alternatively atleast about 98% positives and alternatively at least about 99% positiveswhen compared with the amino acid sequence of a PRO polypeptide having afull-length amino acid sequence as disclosed herein, an amino acidsequence lacking the signal peptide as disclosed herein, anextracellular domain of a transmembrane protein, with or without thesignal peptide, as disclosed herein or any other specifically definedfragment of the full-length amino acid sequence as disclosed herein.

[0488] In a specific aspect, the invention provides an isolated PROpolypeptide without the N-terminal signal sequence and/or the initiatingmethionine and is encoded by a nucleotide sequence that encodes such anamino acid sequence as hereinbefore described. Processes for producingthe same are also herein described, wherein those processes compriseculturing a host cell comprising a vector which comprises theappropriate encoding nucleic acid molecule under conditions suitable forexpression of the PRO polypeptide and recovering the PRO polypeptidefrom the cell culture.

[0489] Another aspect the invention provides an isolated PRO polypeptidewhich is either transmembrane domain-deleted or transmembranedomain-inactivated. Processes for producing the same are also hereindescribed, wherein those processes comprise culturing a host cellcomprising a vector which comprises the appropriate encoding nucleicacid molecule under conditions suitable for expression of the PROpolypeptide and recovering the PRO polypeptide from the cell culture.

[0490] In yet another embodiment, the invention concerns agonists andantagonists of a native PRO polypeptide as defined herein. In aparticular embodiment, the agonist or antagonist is an anti-PRO antibodyor a small molecule.

[0491] In a further embodiment, the invention concerns a method ofidentifying agonists or antagonists to a PRO polypeptide which comprisecontacting tile PRO polypeptide with a candidate molecule and monitoringa biological activity mediated by said PRO polypeptide. Preferably, thePRO polypeptide is a native PRO polypeptide.

[0492] In a still further embodiment, the invention concerns acomposition of matter comprising a PRO polypeptide, or an agonist orantagonist of a PRO polypeptide as herein described, or an anti-PROantibody, in combination with a carrier. Optionally, the carrier is apharmaceutically acceptable carrier.

[0493] Another embodiment of the present invention is directed to theuse of a PRO polypeptide, or an agonist or antagonist thereof ashereinbefore described, or an anti-PRO antibody, for the preparation ofa medicament useful in the treatment of a condition which is responsiveto the PRO polypeptide, an agonist or antagonist thereof or an anti-PROantibody.

BRIEF DESCRIPTION OF THE DRAWINGS

[0494]FIG. 1 shows a nucleotide sequence (SEQ ID NO:1) of a nativesequence PRO1484 cDNA, wherein SEQ ID NO:1 is a clone designated hereinas “DNA44686-1653”.

[0495]FIG. 2 shows the amino acid sequence (SEQ ID NO:2) derived fromthe coding sequence of SEQ ID NO:1 shown in FIG. 1.

[0496]FIG. 3 shows a nucleotide sequence (SEQ ID NO:8) of a nativesequence PRO4334 cDNA, wherein SEQ ID NO:8 is a clone designated hereinas “DNA59608-2577”.

[0497]FIG. 4 shows the amino acid sequence (SEQ ID NO:9) derived fromthe coding sequence of SEQ ID NO:8 shown in FIG. 3.

[0498]FIG. 5 shows a nucleotide sequence (SEQ ID NO:10) of a nativesequence PRO1122 cDNA, wherein SEQ ID NO:10 is a clone designated hereinas “DNA62377-1381”.

[0499]FIG. 6 shows the amino acid sequence (SEQ ID NO:11) derived fromthe coding sequence of SEQ ID NO:10 shown in FIG. 5.

[0500]FIG. 7 shows a nucleotide sequence (SEQ ID NO:15) of a nativesequence PRO1889 cDNA, wherein SEQ ID NO:15 is a clone designated hereinas “DNA77623-2524”.

[0501]FIG. 8 shows the amino acid sequence (SEQ ID NO:16) derived fromthe coding sequence of SEQ ID NO:15 shown in FIG. 7.

[0502]FIG. 9 shows a nucleotide sequence (SEQ ID NO:17) of a nativesequence PRO1890 cDNA, wherein SEQ ID NO:17 is a clone designated hereinas “DNA79230-2525”.

[0503]FIG. 10 shows the amino acid sequence (SEQ ID NO:18) derived fromthe coding sequence of SEQ ID NO:17 shown in FIG. 9.

[0504]FIG. 11 shows a nucleotide sequence (SEQ ID NO:22) of a nativesequence PRO1887 cDNA, wherein SEQ ID NO:22 is a clone designated hereinas “DNA79862-2522”.

[0505]FIG. 12 shows the amino acid sequence (SEQ ID NO:23) derived fromthe coding sequence of SEQ ID NO:22 shown in FIG. 11.

[0506]FIG. 13 shows a nucleotide sequence (SEQ ID NO:28) of a nativesequence PRO1785 cDNA, wherein SEQ ID NO:28 is a clone designated hereinas “DNA80136-2503”.

[0507]FIG. 14 shows the amino acid sequence (SEQ ID NO:29) derived fromthe coding sequence of SEQ ID NO:28 shown in FIG. 13.

[0508]FIG. 15 shows a nucleotide sequence (SEQ ID NO:34) of a nativesequence PRO4353 cDNA, wherein SEQ ID NO:34 is a clone designated hereinas “DNA80145-2594”.

[0509]FIG. 16 shows the amino acid sequence (SEQ ID NO:35) derived fromthe coding sequence of SEQ ID NO:34 shown in FIG. 15.

[0510]FIG. 17 shows a nucleotide sequence (SEQ ID NO:39) of a nativesequence PRO4357 cDNA, wherein SEQ ID NO:39 is a clone designated hereinas “DNA84917-2597”.

[0511]FIG. 18 shows the amino acid sequence (SEQ ID NO:40) derived fromthe coding sequence of SEQ ID NO:39 shown in FIG. 17.

[0512]FIG. 19 shows a nucleotide sequence (SEQ ID NO:44) of a nativesequence PRO4405 cDNA, wherein SEQ ID NO:44 is a clone designated hereinas “DNA84920-2614”.

[0513]FIG. 20 shows the amino acid sequence (SEQ ID NO:45) derived fromthe coding sequence of SEQ ID NO:44 shown in FIG. 19.

[0514]FIG. 21 shows a nucleotide sequence (SEQ ID NO:49) of a nativesequence PRO4356 cDNA, wherein SEQ ID NO:49 is a clone designated hereinas “DNA86576-2595”.

[0515]FIG. 22 shows the amino acid sequence (SEQ ID NO:50) derived fromthe coding sequence of SEQ ID NO:49 shown in FIG. 21.

[0516]FIG. 23 shows a nucleotide sequence (SEQ ID NO:51) of a nativesequence PRO4352 cDNA, wherein SEQ ID NO:51 is a clone designated hereinas “DNA87976-2593”.

[0517]FIG. 24 shows the amino acid sequence (SEQ ID NO:52) derived fromthe coding sequence of SEQ ID NO:51 shown in FIG. 23.

[0518]FIG. 25 shows a nucleotide sequence (SEQ ID NO:56) of a nativesequence PRO4380 cDNA, wherein SEQ ID NO:56 is a clone designated hereinas “DNA92234-2602”.

[0519]FIG. 26 shows the amino acid sequence (SEQ ID NO:57) derived fromthe coding sequence of SEQ ID NO:56 shown in FIG. 25.

[0520]FIG. 27 shows a nucleotide sequence (SEQ ID NO:58) of a nativesequence PRO4354 cDNA, wherein SEQ ID NO:58 is a clone designated hereinas “DNA92256-2596”.

[0521]FIG. 28 shows the amino acid sequence (SEQ ID NO:59) derived fromthe coding sequence of SEQ ID NO:58 shown in FIG. 27.

[0522]FIG. 29 shows a nucleotide sequence (SEQ ID NO:60) of a nativesequence PRO4408 cDNA, wherein SEQ ID NO:60 is a clone designated hereinas “DNA92274-2617”.

[0523]FIG. 30 shows the amino acid sequence (SEQ ID NO:61) derived fromthe coding sequence of SEQ ID NO:60 shown in FIG. 29.

[0524]FIG. 31 shows a nucleotide sequence (SEQ ID NO:62) of a nativesequence PRO5737 cDNA, wherein SEQ ID NO:62 is a clone designated hereinas “DNA92929-2534”.

[0525]FIG. 32 shows the amino acid sequence (SEQ ID NO:63) derived fromthe coding sequence of SEQ ID NO:62 shown in FIG. 31.

[0526]FIG. 33 shows a nucleotide sequence (SEQ ID NO:64) of a nativesequence PRO4425 cDNA, wherein SEQ ID NO:64 is a clone designated hereinas “DNA93011-2637”.

[0527]FIG. 34 shows the amino acid sequence (SEQ ID NO:65) derived fromthe coding sequence of SEQ ID NO:64 shown in FIG. 33.

[0528]FIG. 35 shows a nucleotide sequence (SEQ ID NO:66) of a nativesequence PRO5990 cDNA, wherein SEQ ID NO:66 is a clone designated hereinas “DNA96042-2682”.

[0529]FIG. 36 shows the amino acid sequence (SEQ ID NO:67) derived fromthe coding sequence of SEQ ID NO:66 shown in FIG. 35.

[0530]FIG. 37 shows a nucleotide sequence (SEQ ID NO:71) of a nativesequence PRO6030 cDNA, wherein SEQ ID NO:71 is a clone designated hereinas “DNA96850-2705”.

[0531]FIG. 38 shows the amino acid sequence (SEQ ID NO:72) derived fromthe coding sequence of SEQ ID NO:71 shown in FIG. 37.

[0532]FIG. 39 shows a nucleotide sequence (SEQ ID NO:73) of a nativesequence PRO4424 cDNA, wherein SEQ ID NO:73 is a clone designated hereinas “DNA96857-2636”.

[0533]FIG. 40 shows the amino acid sequence (SEQ ID NO:74) derived fromthe coding sequence of SEQ ID NO:73 shown in FIG. 39.

[0534]FIG. 41 shows a nucleotide sequence (SEQ ID NO:75) of a nativesequence PRO4422 cDNA, wherein SEQ ID NO:75 is a clone designated hereinas “DNA96867-2620”.

[0535]FIG. 42 shows the amino acid sequence (SEQ ID NO:76) derived fromthe coding sequence of SEQ ID NO:75 shown in FIG. 41.

[0536]FIG. 43 shows a nucleotide sequence (SEQ ID NO:77) of a nativesequence PRO4430 cDNA, wherein SEQ ID NO:77 is a clone designated hereinas “DNA96878-2626”.

[0537]FIG. 44 shows the amino acid sequence (SEQ ID NO:78) derived fromthe coding sequence of SEQ ID NO:77 shown in FIG. 43.

[0538]FIG. 45 shows a nucleotide sequence (SEQ ID NO:79) of a nativesequence PRO4499 cDNA, wherein SEQ ID NO:79 is a clone designated hereinas “DNA96889-2641”.

[0539]FIG. 46 shows the amino acid sequence (SEQ ID NO:80) derived fromthe coding sequence of SEQ ID NO:79 shown in FIG. 45.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0540] I. Definitions

[0541] The terms “PRO polypeptide” and “PRO” as used herein and whenimmediately followed by a numerical designation refer to variouspolypeptides, wherein the complete designation (i.e., PRO/number) refersto specific polypeptide sequences as described herein. The terms“PRO/number polypeptide” and “PRO/number” wherein the term “number” isprovided as an actual numerical designation as used herein encompassnative sequence polypeptides and polypeptide variants (which are furtherdefined herein). The PRO polypeptides described herein may be isolatedfrom a variety of sources, such as from human tissue types or fromanother source, or prepared by recombinant or synthetic methods.

[0542] A “native sequence PRO polypeptide” comprises a polypeptidehaving the same amino acid sequence as the corresponding PRO polypeptidederived from nature. Such native sequence PRO polypeptides can beisolated from nature or can be produced by recombinant or syntheticmeans. The term “native sequence PRO polypeptide” specificallyencompasses naturally-occurring truncated or secreted forms of thespecific PRO polypeptide (e.g., an extracellular domain sequence),naturally-occurring variant forms (e.g., alternatively spliced forms)and naturally-occurring allelic variants of the polypeptide. In variousembodiments of the invention, the native sequence PRO polypeptidesdisclosed herein are mature or full-length native sequence polypeptidescomprising the full-length amino acids sequences shown in theaccompanying figures. Start and stop codons are shown in bold font andunderlined in the figures. However, while the PRO polypeptide disclosedin the accompanying figures are shown to begin with methionine residuesdesignated herein as amino acid position 1 in the figures, it isconceivable and possible that other methionine residues located eitherupstream or downstream from the amino acid position 1 in the figures maybe employed as the starting amino acid residue for the PRO polypeptides.

[0543] The PRO polypeptide “extracellular domain” or “ECD” refers to aform of the PRO polypeptide which is essentially free of thetransmembrane and cytoplasmic domains. Ordinarily, a PRO polypeptide ECDwill have less than 1% of such transmembrane and/or cytoplasmic domainsand preferably, will have less than 0.5% of such domains. It will beunderstood that any transmembrane domains identified for the PROpolypeptides of the present invention are identified pursuant tocriteria routinely employed in the art for identifying that type ofhydrophobic domain. The exact boundaries of a transmembrane domain mayvary but most likely by no more than about 5 amino acids at either endof the domain as initially identified herein. Optionally, therefore, anextracellular domain of a PRO polypeptide may contain from about 5 orfewer amino acids on either side of the transmembranedomain/extracellular domain boundary as identified in the Examples orspecification and such polypeptides, with or without the associatedsignal peptide, and nucleic acid encoding them, are comtemplated by thepresent invention.

[0544] The approximate location of the “signal peptides” of the variousPRO polypeptides disclosed herein are shown in the present specificationand/or the accompanying figures. It is noted, however, that theC-terminal boundary of a signal peptide may vary, but most likely by nomore than about 5 amino acids on either side of the signal peptideC-terminal boundary as initially identified herein, wherein theC-terminal boundary of the signal peptide may be identified pursuant tocriteria routinely employed in the art for identifying that type ofamino acid sequence element (e.g., Nielsen et al., Prot. Eng. 10:1-6(1997) and von Heinje et al., Nucl. Acids. Res. 14:46834690 (1986)).Moreover, it is also recognized that, in some cases, cleavage of asignal sequence from a secreted polypeptide is not entirely uniform,resulting in more than one secreted species. These mature polypeptides,where the signal peptide is cleaved within no more than about 5 aminoacids on either side of the C-terminal boundary of the signal peptide asidentified herein, and the polynucleotides encoding them, arecontemplated by the present invention.

[0545] “PRO polypeptide variant” means an active PRO polypeptide asdefined above or below having at least about 80% amino acid sequenceidentity with a full-length native sequence PRO polypeptide sequence asdisclosed herein, a PRO polypeptide sequence lacking the signal peptideas disclosed herein, an extracellular domain of a PRO polypeptide, withor without the signal peptide, as disclosed herein or any other fragmentof a full-length PRO polypeptide sequence as disclosed herein. Such PROpolypeptide variants include, for instance, PRO polypeptides wherein oneor more amino acid residues are added, or deleted, at the N- orC-terminus of the full-length native amino acid sequence. Ordinarily, aPRO polypeptide variant will have at least about 80% amino acid sequenceidentity, alternatively at least about 81% amino acid sequence identity,alternatively at least about 82% amino acid sequence identity,alternatively at least about 83% amino acid sequence identity,alternatively at least about 84% amino acid sequence identity,alternatively at least about 85% amino acid sequence identity,alternatively at least about 86% amino acid sequence identity,alternatively at least about 87% amino acid sequence identity,alternatively at least about 88% amino acid sequence identity,alternatively at least about 89% amino acid sequence identity,alternatively at least about 90% amino acid sequence identity,alternatively at least about 91% amino acid sequence identity,alternatively at least about 92% amino acid sequence identity,alternatively at least about 93% amino acid sequence identity,alternatively at least about 94% amino acid sequence identity,alternatively at least about 95% amino acid sequence identity,alternatively at least about 96% amino acid sequence identity,alternatively at least about 97% amino acid sequence identity,alternatively at least about 98% amino acid sequence identity andalternatively at least about 99% amino acid sequence identity to afull-length native sequence PRO polypeptide sequence as disclosedherein, a PRO polypeptide sequence lacking the signal peptide asdisclosed herein, an extracellular domain of a PRO polypeptide, with orwithout the signal peptide, as disclosed herein or any otherspecifically defined fragment of a full-length PRO polypeptide sequenceas disclosed herein. Ordinarily, PRO variant polypeptides are at leastabout 10 amino acids in length, alternatively at least about 20 aminoacids in length, alternatively at least about 30 amino acids in length,alternatively at least about 40 amino acids in length, alternatively atleast about 50 amino acids in length, alternatively at least about 60amino acids in length, alternatively at least about 70 amino acids inlength, alter natively at least about 80 amino acids in length,alternatively at least about 90 amino acids in length, alternatively atleast about 100 amino acids in length, alternatively at least about 150amino acids in length, alternatively at least about 200 amino acids inlength, alternatively at least about 300 amino acids in length, or more.“Percent (%) amino acid sequence identity” with respect to the PROpolypeptide sequences identified herein is defined as the percentage ofamino acid residues in a candidate sequence that are identical with theamino acid residues in the specific PRO polypeptide sequence, afteraligning the sequences and introducing gaps, if necessary, to achievethe maximum percent sequence identity, and not considering anyconservative substitutions as part of the sequence identity. Alignmentfor purposes of determining percent amino acid sequence identity can beachieved in various ways that are within the skill in the art, forinstance, using publicly available computer software such as BLAST,BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the artcan determine appropriate parameters for measuring alignment, includingany algorithms needed to achieve maximal alignment over the full lengthof the sequences being compared. For purposes herein, however, % aminoacid sequence identity values are generated using the sequencecomparison computer program ALIGN-2, wherein the complete source codefor the ALIGN-2 program is provided in Table 1 below. The ALIGN-2sequence comparison computer program was authored by Genentech, Inc. andthe source code shown in Table 1 below has been filed with userdocumentation in the U.S. Copyright Office, Washington D.C., 20559,where it is registered under U.S. Copyright Registration No. TXU510087.The ALIGN-2 program is publicly available through Genentech, Inc., SouthSan Francisco, Calif. or may be compiled from the source code providedin Table 1 below. The ALIGN-2 program should be compiled for use on aUNIX operating system, preferably digital UNIX V4.0D. All sequencecomparison parameters are set by the ALIGN-2 program and do not vary.

[0546] In situations where ALIGN-2 is employed for amino acid sequencecomparisons, the % amino acid sequence identity of a given amino acidsequence A to, with, or against a given amino acid sequence B (which canalternatively be phrased as a given amino acid sequence A that has orcomprises a certain % amino acid sequence identity to, with, or againsta given amino acid sequence B) is calculated as follows:

100 times the fraction {fraction (X/Y)}

[0547] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program ALIGN-2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A. As examples of % amino acid sequenceidentity calculations using this method, Tables 2 and 3 demonstrate howto calculate the % amino acid sequence identity of the amino acidsequence designated “Comparison Protein” to the amino acid sequencedesignated “PRO”, wherein “PRO” represents the amino acid sequence of ahypothetical PRO polypeptide of interest, “Comparison Protein”represents the amino acid sequence of a polypeptide against which the“PRO” polypeptide of interest is being compared, and “X, “Y” and “Z”each represent different hypothetical amino acid residues.

[0548] Unless specifically stated otherwise, all % amino acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, % aminoacid sequence identity values may also be obtained as described below byusing the WU-BLAST-2 computer program (Altschul et al., Methods inEnzymology 266:460-480 (1996)). Most of the WU-BLAST-2 search parametersare set to the default values. Those not set to default values, i.e.,the adjustable parameters, are set with the following values: overlapspan =1, overlap fraction =0.125, word threshold (T) =11, and scoringmatrix =BLOSUM62. When WU-BLAST-2 is employed, a % amino acid sequenceidentity value is determined by dividing (a) the number of matchingidentical amino acid residues between the amino acid sequence of the PROpolypeptide of interest having a sequence derived from the native PROpolypeptide and the comparison amino acid sequence of interest (i.e.,the sequence against which the PRO polypeptide of interest is beingcompared which may be a PRO variant polypeptide) as determined byWU-BLAST-2 by (b) the total number of amino acid residues of the PROpolypeptide of interest. For example, in the statement “a polypeptidecomprising an the amino acid sequence A which has or having at least 80%amino acid sequence identity to the amino acid sequence B”, the aminoacid sequence A is the comparison amino acid sequence of interest andthe amino acid sequence B is the amino acid sequence of the PROpolypeptide of interest.

[0549] Percent amino acid sequence identity may also be determined usingthe sequence comparison program NCBI-BLAST2 (Altschul et al., NucleicAcids Res. 25:3389-3402 (1997)). The NCBI-BLAST2 sequence comparisonprogram may be downloaded from http://www.ncbi.nlm.nih.gov or otherwiseobtained from the National Institute of Health, Bethesda, Md.NCBI-BLAST2 uses several search parameters, wherein all of those searchparameters are set to default values including, for example, unmask=yes, strand =all, expected occurrences =10, minimum low complexitylength =15/5, multi-pass e-value =0.01, constant for multi-pass =25,dropoff for final gapped alignment =25 and scoring matrix =BLOSUM62.

[0550] In situations where NCBI-BLAST2 is employed for amino acidsequence comparisons, the % amino acid sequence identity of a givenamino acid sequence A to, with, or against a given amino acid sequence B(which can alternatively be phrased as a given amino acid sequence Athat has or comprises a certain % amino acid sequence identity to, with,or against a given amino acid sequence B) is calculated as follows:

100 times the fraction {fraction (X/Y)}

[0551] where X is the number of amino acid residues scored as identicalmatches by the sequence alignment program NCBI-BLAST2 in that program'salignment of A and B, and where Y is the total number of amino acidresidues in B. It will be appreciated that where the length of aminoacid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not equal the % amino acidsequence identity of B to A.

[0552] “PRO variant polynucleotide” or “PRO variant nucleic acidsequence” means a nucleic acid molecule which encodes an active PROpolypeptide as defined below and which has at least about 80% nucleicacid sequence identity with a nucleotide acid sequence encoding afull-length native sequence PRO polypeptide sequence as disclosedherein, a full-length native sequence PRO polypeptide sequence lackingthe signal peptide as disclosed herein, an extracellular domain of a PROpolypeptide, with or without the signal peptide, as disclosed herein orany other fragment of a full-length PRO polypeptide sequence asdisclosed herein. Ordinarily, a PRO variant polynucleotide will have atleast about 80% nucleic acid sequence identity, alternatively at leastabout 81% nucleic acid sequence identity, alternatively at least about82% nucleic acid sequence identity, alternatively at least about 83%nucleic acid sequence identity, alternatively at least about 84% nucleicacid sequence identity, alternatively at least about 85% nucleic acidsequence identity, alternatively at least about 86% nucleic acidsequence identity, alternatively at least about 87% nucleic acidsequence identity, alternatively at least about 88% nucleic acidsequence identity, alternatively at least about 89% nucleic acidsequence identity, alternatively at least about 90% nucleic acidsequence identity, alternatively at least about 91% nucleic acidsequence identity, alternatively at least about 92% nucleic acidsequence identity, alternatively at least about 93% nucleic acidsequence identity, alternatively at least about 94% nucleic acidsequence identity, alternatively at least about 95% nucleic acidsequence identity, alternatively at least about 96% nucleic acidsequence identity, alternatively at least about 97% nucleic acidsequence identity, alternatively at least about 98% nucleic acidsequence identity and alternatively at least about 99% nucleic acidsequence identity with a nucleic acid sequence encoding a full-lengthnative sequence PRO polypeptide sequence as disclosed herein, afill-length native sequence PRO polypeptide sequence lacking the signalpeptide as disclosed herein, an extracellular domain of a PROpolypeptide, with or without the signal sequence, as disclosed herein orany other fragment of a full-length PRO polypeptide sequence asdisclosed herein. Variants do not encompass the native nucleotidesequence.

[0553] Ordinarily, PRO variant polynucleotides are at least about 30nucleotides in length, alternatively at least about 60 nucleotides inlength, alternatively at least about 90 nucleotides in length,alternatively at least about 120 nucleotides in length, alternatively atleast about 150 nucleotides in length, alternatively at least about 180nucleotides in length, alternatively at least about 210 nucleotides inlength, alternatively at least about 240 nucleotides in length,alternatively at least about 270 nucleotides in length, alternatively atleast about 300 nucleotides in length, alternatively at least about 450nucleotides in length, alternatively at least about 600 nucleotides inlength, alternatively at least about 900 nucleotides in length, or more.

[0554] “Percent (%) nucleic acid sequence identity” with respect toPRO-encoding nucleic acid sequences identified herein is defined as thepercentage of nucleotides in a candidate sequence that are identicalwith the nucleotides in the PRO nucleic acid sequence of interest, afteraligning the sequences and introducing gaps, if necessary, to achievethe maximum percent sequence identity. Alignment for purposes ofdetermining percent nucleic acid sequence identity can be achieved invarious ways that are within the skill in the art, for instance, usingpublicly available computer software such as BLAST, BLAST-2, ALIGN orMegalign (DNASTAR) software. For purposes herein, however, % nucleicacid sequence identity values are generated using the sequencecomparison computer program ALIGN-2, wherein the complete source codefor the ALIGN-2 program is provided in Table 1 below. The ALIGN-2sequence comparison computer program was authored by Genentech, Inc. andthe source code shown in Table 1 below has been filed with userdocumentation in the U.S. Copyright Office, Washington D.C., 20559,where it is registered under U.S. Copyright Registration No. TXU510087.The ALIGN-2 program is publicly available through Genentech, Inc., SouthSan Francisco, Calif. or may be compiled from the source code providedin Table 1 below. The ALIGN-2 program should be compiled for use on aUNIX operating system, preferably digital UNIX V4.OD. All sequencecomparison parameters are set by the ALIGN-2 program and do not vary.

[0555] In situations where ALIGN-2 is employed for nucleic acid sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction {fraction (W/Z)}

[0556] where W is the number of nucleotides scored as identical matchesby the sequence alignment program ALIGN-2 in that program's alignment ofC and D, and where Z is the total number of nucleotides in D. It will beappreciated that where the length of nucleic acid sequence C is notequal to the length of nucleic acid sequence D, the % nucleic acidsequence identity of C to D will not equal the % nucleic acid sequenceidentity of D to C. As examples of % nucleic acid sequence identitycalculations, Tables 4 and 5, demonstrate how to calculate the % nucleicacid sequence identity of the nucleic acid sequence designated“Comparison DNA” to the nucleic acid sequence designated “PRO-DNA”,wherein “PRO-DNA” represents a hypothetical PRO-encoding nucleic acidsequence of interest, “Comparison DNA” represents the nucleotidesequence of a nucleic acid molecule against which the “PRO-DNA” nucleicacid molecule of interest is being compared, and “N”, “L” and “V” eachrepresent different hypothetical nucleotides.

[0557] Unless specifically stated otherwise, all % nucleic acid sequenceidentity values used herein are obtained as described in the immediatelypreceding paragraph using the ALIGN-2 computer program. However, %nucleic acid sequence identity values may also be obtained as describedbelow by using the WU-BLAST-2 computer program (Altschul et al., Methodsin Enzymology 266:460-480 (1996)). Most of the WU-BLAST-2 searchparameters are set to the default values. Those not set to defaultvalues, i.e., the adjustable parameters, are set with the followingvalues: overlap span =1, overlap fraction =0.125, word threshold (T)=11, and scoring matrix =BLOSUM62. When WU-BLAST-2 is employed, a %nucleic acid sequence identity value is determined by dividing (a) thenumber of matching identical nucleotides between the nucleic acidsequence of the PRO polypeptide-encoding nucleic acid molecule ofinterest having a sequence derived from the native sequence PROpolypeptide-encoding nucleic acid and the comparison nucleic acidmolecule of interest (i.e., the sequence against which the PROpolypeptide-encoding nucleic acid molecule of interest is being comparedwhich may be a variant PRO polynucleotide) as determined by WU-BLAST-2by (b) the total number of nucleotides of the PRO polypeptide-encodingnucleic acid molecule of interest. For example, in the statement “anisolated nucleic acid molecule comprising a nucleic acid sequence Awhich has or having at least 80% nucleic acid sequence identity to thenucleic acid sequence B”, the nucleic acid sequence A is the comparisonnucleic acid molecule of interest and the nucleic acid sequence B is thenucleic acid sequence of the PRO polypeptide-encoding nucleic acidmolecule of interest.

[0558] Percent nucleic acid sequence identity may also be determinedusing the sequence comparison program NCBI-BLAST2 (Altschul et al.,Nucleic Acids Res. 25:3389-3402 (1997)). The NCBI-BLAST2 sequencecomparison program may be downloaded from http://www.ncbi.nlm.nih.gov orotherwise obtained from the National Institute of Health, Bethesda, MD.NCBI-BLAST2 uses several search parameters, wherein all of those searchparameters are set to default values including, for example, unmask=yes, strand =all, expected occurrences =10, minimum low complexitylength =15/5, multi-pass e-value =0.01, constant for multi-pass =25,dropoff for final gapped alignment =25 and scoring matrix =BLOSUM62.

[0559] In situations where NCBI-BLAST2 is employed for sequencecomparisons, the % nucleic acid sequence identity of a given nucleicacid sequence C to, with, or against a given nucleic acid sequence D(which can alternatively be phrased as a given nucleic acid sequence Cthat has or comprises a certain % nucleic acid sequence identity to,with, or against a given nucleic acid sequence D) is calculated asfollows:

100 times the fraction {fraction (W/Z)}

[0560] where W is the number of nucleotides scored as identical matchesby the sequence alignment program NCBI-BLAST2 in that program'salignment of C and D, and where Z is the total number of nucleotides inD. It will be appreciated that where the length of nucleic acid sequenceC is not equal to the length of nucleic acid sequence D, the % nucleicacid sequence identity of C to D will not equal the % nucleic acidsequence identity of D to C.

[0561] In other embodiments, PRO variant polynucleotides are nucleicacid molecules that encode an active PRO polypeptide and which arecapable of hybridizing, preferably under stringent hybridization andwash conditions, to nucleotide sequences encoding a full-length PROpolypeptide as disclosed herein. PRO variant polypeptides may be thosethat are encoded by a PRO variant polynucleotide.

[0562] The term “positives”, in the context of sequence comparisonperformed as described above, includes residues in the sequencescompared that are not identical but have similar properties (e.g. as aresult of conservative substitutions, see Table 6 below). For purposesherein, the % value of positives is determined by dividing (a) thenumber of amino acid residues scoring a positive value between the PROpolypeptide amino acid sequence of interest having a sequence derivedfrom the native PRO polypeptide sequence and the comparison amino acidsequence of interest (i.e., the amino acid sequence against which thePRO polypeptide sequence is being compared) as determined in theBLOSUM62 matrix of WU-BLAST-2 by (b) the total number of amino acidresidues of the PRO polypeptide of interest.

[0563] Unless specifically stated otherwise, the % value of positives iscalculated as described in the immediately preceding paragraph. However,in the context of the amino acid sequence identity comparisons performedas described for ALIGN-2 and NCBI-BLAST-2 above, includes amino acidresidues in the sequences compared that are not only identical, but alsothose that have similar properties. Amino acid residues that score apositive value to an amino acid residue of interest are those that areeither identical to the amino acid residue of interest or are apreferred substitution (as defined in Table 6 below) of the amino acidresidue of interest.

[0564] For amino acid sequence comparisons using ALIGN-2 or NCBI-BLAST2,the % value of positives of a given amino acid sequence A to, with, oragainst a given amino acid sequence B (which can alternatively bephrased as a given amino acid sequence A that has or comprises a certain% positives to, with, or against a given amino acid sequence B) iscalculated as follows:

100 times the fraction {fraction (X/Y)}

[0565] where X is the number of amino acid residues scoring a positivevalue as defined above by the sequence alignment program ALIGN-2 orNCBI-BLAST2 in that program's alignment of A and B, and where Y is thetotal number of amino acid residues in B. It will be appreciated thatwhere the length of amino acid sequence A is not equal to the length ofamino acid sequence B, the % positives of A to B will not equal the %positives of B to A.

[0566] “Isolated,” when used to describe the various polypeptidesdisclosed herein, means polypeptide that has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials thatwould typically interfere with diagnostic or therapeutic uses for thepolypeptide, and may include enzymes, hormones, and other proteinaceousor non-proteinaceous solutes. In preferred embodiments, the polypeptidewill be purified (1) to a degree sufficient to obtain at least 15residues of N-terminal or internal amino acid sequence by use of aspinning cup sequenator, or (2) to homogeneity by SDS-PAGE undernon-reducing or reducing conditions using Coomassie blue or, preferably,silver stain. Isolated polypeptide includes polypeptide in situ withinrecombinant cells, since at least one component of the PRO polypeptidenatural environment will not be present. Ordinarily, however, isolatedpolypeptide will be prepared by at least one purification step.

[0567] An “isolated” PRO polypeptide-encoding nucleic acid or otherpolypeptide-encoding nucleic acid is a nucleic acid molecule that isidentified and separated from at least one contaminant nucleic acidmolecule with which it is ordinarily associated in the natural source ofthe polypeptide-encoding nucleic acid. An isolated polypeptide-encodingnucleic acid molecule is other than in the form or setting in which itis found in nature. Isolated polypeptide-encoding nucleic acid moleculestherefore are distinguished from the specific polypeptide-encodingnucleic acid molecule as it exists in natural cells. However, anisolated polypeptide-encoding nucleic acid molecule includespolypeptide-encoding nucleic acid molecules contained in cells thatordinarily express the polypeptide where, for example, the nucleic acidmolecule is in a chromosomal location different from that of naturalcells.

[0568] The term “control sequences” refers to DNA sequences necessaryfor the expression of an operably linked coding sequence in a particularhost organism. The control sequences that are suitable for prokaryotes,for example, include a promoter, optionally an operator sequence, and aribosome binding site. Eukaryotic cells are known to utilize promoters,polyadenylation signals, and enhancers.

[0569] Nucleic acid is “operably linked” when it is placed into afunctional relationship with another nucleic acid sequence. For example,DNA for a presequence or secretory leader is operably linked to DNA fora polypeptide if it is expressed as a preprotein that participates inthe secretion of the polypeptide; a promoter or enhancer is operablylinked to a coding sequence if it affects the transcription of thesequence; or a ribosome binding site is operably linked to a codingsequence if it is positioned so as to facilitate translation. Generally,operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading phase. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites. If suchsites do not exist, the synthetic oligonucleotide adaptors or linkersare used in accordance with conventional practice.

[0570] The term “antibody” is used in the broadest sense andspecifically covers, for example, single anti-PRO monoclonal antibodies(including agonist, antagonist, and neutralizing antibodies), anti-PROantibody compositions with polyepitopic specificity, single chainanti-PRO antibodies, and fragments of anti-PRO antibodies (see below).The term “monoclonal antibody” as used herein refers to an antibodyobtained from a population of substaniially homogeneous antibodies,i.e., the individual antibodies comprising the population are identicalexcept for possible naturally-occurring mutations that may be present inminor amounts.

[0571] “Stringency” of hybridization reactions is readily determinableby one of ordinary skill in the art, and generally is an empiricalcalculation dependent upon probe length, washing temperature, and saltconcentration. In general, longer probes require higher temperatures forproper annealing, while shorter probes need lower temperatures.Hybridization generally depends on the ability of denatured DNA toreanneal when complementary strands are present in an environment belowtheir melting temperature. The higher the degree of desired homologybetween the probe and hybridizable sequence, the higher the relativetemperature which can be used. As a result, it follows that higherrelative temperatures would tend to make the reaction conditions morestringent, while lower temperatures less so. For additional details andexplanation of stringency of hybridization reactions, see Ausubel etal., Current Protocols in Molecular Biology, Wiley IntersciencePublishers, (1995).

[0572] “Stringent conditions” or “high stringency conditions”, asdefined herein, may be identified by those that: (1) employ low ionicstrength and high temperature for washing, for example 0.015 M sodiumchloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.;(2) employ during hybridization a denaturing agent, such as formamide,for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1%Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; or (3)employ 50% formamide, 5× SSC (0.75 M NaCl, 0.075 M sodium citrate), 50mnM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5× Derhardt'ssolution, sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10%dextran sulfate at 42° C., with washes at 42° C. in 0.2× SSC (sodiumchloride/sodium citrate) and 50% formamide at 55° C., followed by ahigh-stringency wash consisting of 0.1× SSC containing EDTA at 55° C.

[0573] “Moderately stringent conditions” may be identified as describedby Sambrook et al., Molecular Cloning: A Laboratoiy Manual, New York:Cold Spring Harbor Press, 1989, and include the use of washing solutionand hybridization conditions (e.g., temperature, ionic strength and%SDS) less stringent that those described above. An example ofmoderately stringent conditions is overnight incubation at 37° C. in asolution comprising: 20% formamide, 5× SSC (150 mM NaCl, 15 mM trisodiumcitrate), 50 mM sodium phosphate (pH 7.6), 5× Denhardt's solution, 10%dextran sulfate, and 20 mg/ml denatured sheared salmon sperm DNA,followed by washing the filters in 1× SSC at about 37-50° C. The skilledartisan will recognize how to adjust the temperature, ionic strength,etc. as necessary to accommodate factors such as probe length and thelike.

[0574] The term “epitope tagged” when used herein refers to a chimericpolypeptide comprising a PRO polypeptide fused to a “tag polypeptide”.The tag polypeptide has enough residues to provide an epitope againstwhich an antibody can be made, yet is short enough such that it does notinterfere with activity of the polypeptide to which it is fused. The tagpolypeptide preferably also is fairly unique so that the antibody doesnot substantially cross-react with other epitopes. Suitable tagpolypeptides generally have at least six amino acid residues and usuallybetween about 8 and 50 amino acid residues (preferably, between about 10and 20 amino acid residues).

[0575] As used herein, the term “immunoadhesin” designates antibody-likemolecules which combine the binding specificity of a heterologousprotein (an “adhesin”) with the effector functions of immunoglobulinconstant domains. Structurally, the immunoadhesins comprise a fusion ofan amino acid sequence with the desired binding specificity which isother than the antigen recognition and binding site of an antibody(i.e., is “heterologous”), and an immunoglobulin constant domainsequence. The adhesin part of an immunoadhesin molecule typically is acontiguous amino acid sequence comprising at least the binding site of areceptor or a ligand. The immunoglobulin constant domain sequence in theimmunoadhesin may be obtained from any immunoglobulin, such as IgG-1,IgG-2, IgG-3, or IgG4 subtypes, IgA (including IgA-1 and IgA-2), IgE,IgD or IgM.

[0576] “Active” or “activity” for the purposes herein refers to form(s)of a PRO polypeptide which retain a biological and/or an immunologicalactivity of native or naturally-occurring PRO, wherein “biological”activity refers to a biological function (either inhibitory orstimulatory) caused by a native or naturally-occurring PRO other thanthe ability to induce the production of an antibody against an antigenicepitope possessed by a native or naturally-occurring PRO and an“immunological” activity refers to the ability to induce the productionof an antibody against an antigenic epitope possessed by a native ornaturally-occurring PRO.

[0577] The term “antagonist” is used in the broadest sense, and includesany molecule that partially or fully blocks, inhibits, or neutralizes abiological activity of a native PRO polypeptide disclosed herein. In asimilar manner, the term “agonist” is used in the broadest sense andincludes any molecule that mimics a biological activity of a native PROpolypeptide disclosed herein. Suitable agonist or antagonist moleculesspecifically include agonist or antagonist antibodies or antibodyfragments, fragments or amino acid sequence variants of native PROpolypeptides, peptides, antisense oligonucleotides, small organicmolecules, etc. Methods for identifying agonists or antagonists of a PROpolypeptide may comprise contacting a PRO polypeptide with a candidateagonist or antagonist molecule and measuring a detectable change in oneor more biological activities normally associated with the PROpolypeptide.

[0578] “Treatment” refers to both therapeutic treatment and prophylacticor preventative measures, wherein the object is to prevent or slow down(lessen) the targeted pathologic condition or disorder. Those in need oftreatment include those already with the disorder as well as those proneto have the disorder or those in whom the disorder is to be prevented.

[0579] “Chronic” administration refers to administration of the agent(s)in a continuous mode as opposed to an acute mode, so as to maintain theinitial therapeutic effect (activity) for an extended period of time.“Intermittent” administration is treatment that is not consecutivelydone without interruption, but rather is cyclic in nature.

[0580] “Mammal” for purposes of treatment refers to any animalclassified as a mammal, including humans, domestic and farm animals, andzoo, sports, or pet animals, such as dogs, cats, cattle, horses, sheep,pigs, goats, rabbits, etc. Preferably, the mammal is human.

[0581] Administration “in combination with” one or more furthertherapeutic agents includes simultaneous (concurrent) and consecutiveadministration in any order.

[0582] “Carriers” as used herein include pharmaceutically acceptablecarriers, excipients, or stabilizers which are nontoxic to the cell ormammal being exposed thereto at the dosages and concentrations employed.Often the physiologically acceptable carrier is an aqueous pH bufferedsolution. Examples of physiologically acceptable carriers includebuffers such as phosphate, citrate, and other organic acids;antioxidants including ascorbic acid; low molecular weight (less thanabout 10 residues) polypeptide; proteins, such as serum albumin,gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mannose, or dextrins; chelatingagents such as EDTA; sugar alcohols such as mannitol or sorbitol;salt-forming counterions such as sodium; and/or nonionic surfactantssuch as TWEEN™, polyethylene glycol (PEG), and PLURONICS™. “Antibodyfragments” comprise a portion of an intact antibody, preferably theantigen binding or variable region of the intact antibody. Examples ofantibody fragments include Fab, Fab′, F(ab′)₂, and Fv fragments;diabodies; linear antibodies (Zapata et al., Protein Eng. 8(10):1057-1062 [1995]); single-chain antibody molecules; and multispecificantibodies formed from antibody fragments.

[0583] Papain digestion of antibodies produces two identicalantigen-binding fragments, called “Fab” fragments, each with a singleantigen-binding site, and a residual “Fc” fragment, a designationreflecting the ability to crystallize readily. Pepsin treatment yieldsan F(ab′)₂ fragment that has two antigen-combining sites and is stillcapable of cross-linking antigen.

[0584] “Fv” is the minimum antibody fragment which contains a completeantigen-recognition and -binding site. This region consists of a dimerof one heavy- and one light-chain variable domain in tight, non-covalentassociation. It is in this configuration that the three CDRs of eachvariable domain interact to define an antigen-binding site on thesurface of the V_(H)-V_(L) dimer. Collectively, the six CDRs conferantigen-binding specificity to the antibody. However, even a singlevariable domain (or half of an Fv comprising only three CDRs specificfor an antigen) has the ability to recognize and bind antigen, althoughat a lower affinity than the entire binding site.

[0585] The Fab fragment also contains the constant domain of the lightchain and the first constant domain (CHI) of the heavy chain. Fabfragments differ from Fab′ fragments by the addition of a few residuesat the carboxy terminus of the heavy chain CH1 domain including one ormore cysteines from the antibody hinge region. Fab′-SH is thedesignation herein for Fab′ in which the cysteine residue(s) of theconstant domains bear a free thiol group. F(ab′)₂ antibody fragmentsoriginally were produced as pairs of Fab′ fragments which have hingecysteines between them. Other chemical couplings of antibody fragmentsare also known.

[0586] The “light chains” of antibodies (immunoglobulins) from anyvertebrate species can be assigned to one of two clearly distinct types,called kappa and lambda, based on the amino acid sequences of theirconstant domains.

[0587] Depending on the amino acid sequence of the constant domain oftheir heavy chains, immunoglobulins can be assigned to differentclasses. There are five major classes of immunoglobulins: IgA, IgD, IgE,IgG, and IgM, and several of these may be further divided intosubclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2.

[0588] “Single-chain Fv” or “sFv” antibody fragments comprise the V_(H)and V_(L) domains of antibody, wherein these domains are present in asingle polypeptide chain. Preferably, the Fv polypeptide furthercomprises a polypeptide linker between the V_(H) and V_(L) domains whichenables the sFv to form the desired structure for antigen binding. For areview of sFv, see Pluckthum in The Pharmacology of MonoclonalAntibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, NewYork, pp. 269-315 (1994).

[0589] The term “diabodies” refers to small antibody fragments with twoantigen-binding sites, which fragments comprise a heavy-chain variabledomain (V_(H)) connected to a light-chain variable domain (V_(L)) in thesame polypeptide chain (V_(H)-V_(L)). By using a linker that is tooshort to allow pairing between the two domains on the same chain, thedomains are forced to pair with the complementary domains of anotherchain and create two antigen-binding sites. Diabodies are described morefully in, for example, EP 404,097; WO 93/11161; and Hollinger et al.,Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).

[0590] An “isolated” antibody is one which has been identified andseparated and/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials whichwould interfere with diagnostic or therapeutic uses for the antibody,and may include enzymes, hormones, and other proteinaceous ornonproteinaceous solutes. In preferred embodiments, the antibody will bepurified (1) to greater than 95% by weight of antibody as determined bythe Lowry method, and most preferably more than 99% by weight, (2) to adegree sufficient to obtain at least 15 residues of N-terminal orinternal amino acid sequence by use of a spinning cup sequenator, or (3)to homogeneity by SDS-PAGE under reducing or nonreducing conditionsusing Coomassie blue or, preferably, silver stain. Isolated antibodyincludes the antibody in situ within recombinant cells since at leastone component of the antibody's natural environment will not be present.Ordinarily, however, isolated antibody will be prepared by at least onepurification step.

[0591] The word “label” when used herein refers to a detectable compoundor composition which is conjugated directly or indirectly to theantibody so as to generate a “labeled” antibody. The label may bedetectable by itself (e.g. radioisotope labels or fluorescent labels)or, in the case of an enzymatic label, may catalyze chemical alterationof a substrate compound or composition which is detectable.

[0592] By “solid phase” is meant a non-aqueous matrix to which theantibody of the present invention can adhere. Examples of solid phasesencompassed herein include those formed partially or entirely of glass(e.g., controlled pore glass), polysaccharides (e.g., agarose),polyacrylamides, polystyrene, polyvinyl alcohol and silicones. Incertain embodiments, depending on the context, the solid phase cancomprise the well of an assay plate; in others it is a purificationcolumn (e.g., an affinity chromatography column). This term alsoincludes a discontinuous solid phase of discrete particles, such asthose described in U.S. Pat. No. 4,275,149.

[0593] A “liposome” is a small vesicle composed of various types oflipids, phospholipids and/or surfactant which is useful for delivery ofa drug (such as a PRO polypeptide or antibody thereto) to a mammal. Thecomponents of the liposome are commonly arranged in a bilayer formation,similar to the lipid arrangement of biological membranes.

[0594] A “small molecule” is defined herein to have a molecular weightbelow about 500 Daltons.

TABLE 2 PRO XXXXXXXXXXXXXXX (Length = 15 amino acids) ComparisonXXXXXYYYYYYY (Length = 12 amino acids) Protein

[0595] TABLE 3 PRO XXXXXXXXXX (Length = 10 amino acids) ComparisonXXXXXYYYYYYZZYZ (Length = 15 amino acids) Protein

[0596] TABLE 4 PRO-DNA NNNNNNNNNNNNNN (Length = 14 nucleotides)Comparison NNNNNNLLLLLLLLLL (Length = 16 nucleotides) DNA

[0597] TABLE 5 PRO-DNA NNNNNNNNNNNN (Length = 12 nucleotides) ComparisonNNNNLLLVV (Length = 9 nucleotides) DNA

[0598] II. Compositions and Methods of the Invention

[0599] A. Full-Length PRO Polypeptides

[0600] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO polypeptides. In particular, cDNAs encoding variousPRO polypeptides have been identified and isolated, as disclosed infurther detail in the Examples below. It is noted that proteins producedin separate expression rounds may be given different PRO numbers but theUNQ number is unique for any given DNA and the encoded protein, and willnot be changed. However, for sake of simplicity, in the presentspecification the protein encoded by the full length native nucleic acidmolecules disclosed herein as well as all further native homologues andvariants included in the foregoing definition of PRO, will be referredto as “PRO/number”, regardless of their origin or mode of preparation.

[0601] As disclosed in the Examples below, various cDNA clones have beendeposited with the ATCC. The actual nucleotide sequences of those clonescan readily be determined by the skilled artisan by sequencing of thedeposited clone using routine methods in the art. The predicted aminoacid sequence can be determined from the nucleotide sequence usingroutine skill. For the PRO polypeptides and encoding nucleic acidsdescribed herein, Applicants have identified what is believed to be thereading frame best identifiable with the sequence information availableat the time.

[0602] 1. Full-length PRO1484 Polypeptides

[0603] Using the WU-BLAST2 sequence alignment computer program, it hasbeen found that a full-length native sequence PRO1484 (shown in FIG. 2and SEQ ID NO:2) has certain amino acid sequence identity with a portionof the mouse adipocyte complement related protein (ACR3_MOUSE).Accordingly, it is presently believed that PRO1484 disclosed in thepresent application is a newly identified adipocyte complement-relatedprotein homolog and may possess activity typical of that protein.

[0604] 2. Full-length PRO4334 Polypeptides

[0605] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4334 (shown in FIG. 4 andSEQ ID NO:9) has certain amino acid sequence identity with PC-1.Accordingly, it is presently believed that PRO4334 disclosed in thepresent application is a newly identified member of the PC-1 family andshares similar mechanisms.

[0606] 3. Full-length PRO1122 Polypeptides

[0607] The present invention provides newly identified and isolatednucleotide sequences encoding polypeptides referred to in the presentapplication as PRO1122. In particular, Applicants have identified andisolated cDNA encoding a PRO1122 polypeptide, as disclosed in furtherdetail in the Examples below. Using BLAST and FastA sequence alignmentcomputer programs, Applicants found that the PRO1122 polypeptide hassequence identity with CTLA-8. The amino acid sequence shows a regionhaving sequence identity with IL-17. Accordingly, it is presentlybelieved that PRO1122 polypeptide disclosed in the present applicationis a novel cytokine and thus may be involved in inflammation responses.

[0608] 4. Full-length PRO1889 Polypeptides

[0609] Using the WU-BLAST2 sequence alignment computer program, it hasbeen found that a portion of the full-length native sequence PRO1889(shown in FIG. 8 and SEQ ID NO:16) has certain amino acid sequenceidentity with a portion of the human E48 antigen protein(HSE48ATGN_(—)1). Accordingly, it is presently believed that PRO1889disclosed in the present application is a newly identified E48 homologand may possess activity or properties typical of the E48 protein.

[0610] 5. Full-length PRO1890 Polypeptides

[0611] Using the WU-BLAST2 sequence alignment computer program, it hasbeen found that a portion of the full-length native sequence PRO1890(shown in FIG. 10 and SEQ ID NO:18) has certain amino acid sequenceidentity with a portion of the layilin protein (AF093673_(—)1).

[0612] 6. Full-length PRO1887 Polypeptides

[0613] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO1887 (FIG. 12; SEQ ID NO:23)has certain amino acid sequence identity with a mouse livercarboxylesterase precursor identified on the Dayhoff database as“ESTM_MOUSE”. Accordingly, it is presently believed that PRO1887disclosed in the present application is a newly identified member of thecarboxylesterase family and may possess enzymatic activity typical ofcarboxylesterases.

[0614] 7. Full-length PRO1785 Polypeptides

[0615] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO1785 (shown in FIG. 14 andSEQ ID NO:29) has certain amino acid sequence identity with glutathioneperoxidase. Accordingly, it is presently believed that PRO1785 disclosedin the present application is a newly identified member of theperoxidase family and may possess antioxidant enzyme activity.Regulation of antioxidant activity is of interest in the treatment ofcancer and aging.

[0616] 8. Full-length PRO4353 Polypeptides

[0617] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4353 (shown in FIG. 16 andSEQ ID NO:35) has certain amino acid sequence identity with semaphorinZ. Accordingly, it is presently believed that PRO4353 disclosed in thepresent application is a newly identified member of the semaphorin Zfamily and is involved in inhibition of nerve growth. PRO4353 can beused in assays to identify modulators of semaphorin Z, particularlyinhibitors to promote central nerve regeneration.

[0618] 9. Full-length PRO4357 Polypeptides

[0619] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4357 (shown in FIG. 18 andSEQ ID NO:40) has certain amino acid sequence identity with 289 aminoacids in accession number P_W48804.However, PRO4357 has an additional213 amino acids at the N-terminus.

[0620] 10. Full-length PRO4405 Polypeptides

[0621] As far as is known, the DNA84920-2614 sequence encodes a novelfactor designated herein as PRO4405; using WU-BLAST2 sequence alignmentcomputer programs, limited sequence identities to known proteins wererevealed.

[0622] 11. Full-length PRO4356 Polypeptides

[0623] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4356 (shown in FIG. 22 andSEQ ID NO:50) has certain amino acid sequence identity with metastasisassociated GPI-anchored protein. Accordingly, it is presently believedthat PRO4356 disclosed in the present application is a newly identifiedmember of this family and shares similar mechanisms.

[0624] 12. Full-length PRO4352 Polypeptides

[0625] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4352 (shown in FIG. 24 andSEQ ID NO:52) has certain amino acid sequence identity withprotocadherin pc3 and protocadherin pc4. Accordingly, it is believedthat PRO4352 is involved in cell adhesin and can be used in treatmentsregarding differentiation disorders, cell adhesin, neural receptor orskin disorders. Moreover, it can be used in screens to identify agonistsand antagonists to treat such disorders.

[0626] 13. Full-length PRO4380 Polypeptides

[0627] As far as is known, the DNA92234-2602 sequence encodes a novelfactor designated herein as PRO4380; using WU-BLAST2 sequence alignmentcomputer programs, limited sequence identities to proteins with knownfunctions were revealed.

[0628] 14. Full-length PRO4354 Polypeptides

[0629] As far as is known, the DNA92256-2596 sequence encodes a novelfactor designated herein as PRO4354; using WU-BLAST2 sequence alignmentcomputer programs, limited sequence identities to proteins with knownfunctions were revealed.

[0630] 15. Full-length PRO4408 Polypeptides

[0631] As far as is known, the DNA92274-2617 sequence encodes a novelfactor designated herein as PRO4408; using WU-BLAST2 sequence alignmentcomputer programs, limited sequence identities to known proteins wererevealed.

[0632] 16. Full-length PRO5737 Polypeptides

[0633] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO5737 (shown in FIG. 32 andSEQ ID NO:63) has certain amino acid sequence identity with IL-1 and/orIL-1Ra. Accordingly, it is presently believed that PRO5737 disclosed inthe present application is a newly identified member of this family andshares similar mechanisms.

[0634] 17. Full-length PRO4425 Polypeptides

[0635] As far as is known, the DNA93011-2637 sequence encodes a novelfactor designated herein as PRO4425; using WU-BLAST2 sequence alignmentcomputer programs, PRO4425 showed homology to a protein in GenBank,accession number HGS_RE295, but is not identical.

[0636] 18. Full-length PRO5990 Polypeptides

[0637] Using the ALIGN-2 sequence alignment computer program referencedabove, it has been found that the full-length native sequence PRO5990(shown in FIG. 36 and SEQ ID NO:67) has certain amino acid sequenceidentity with Secretogranin II (Dayhoff No. GEN14673). Accordingly, itis presently believed that the PRO5990 polypeptide disclosed in thepresent application is a newly identified member of the secretograninprotein family and may possess one or more biological and/orimmunological activities or properties typical of that protein family.

[0638] 19. Full-length PRO6030 Polypeptides

[0639] The DNA96850-2705 clone was isolated from a human library asdescribed in the Examples below. As far as is known, the DNA96850-2705nucleotide sequence encodes a novel factor designated herein as PRO6030;using the AI,IGN-2 sequence alignment computer program, no significantsequence identities to any known proteins were revealed.

[0640] 20. Full-length PRO4424 Polypeptides

[0641] As far as is known, the DNA96857-2636 sequence encodes a novelfactor designated herein as PRO4424; using WU-BLAST2 sequence alignmentcomputer programs, PRO4424 showed homology to a protein in GenBank,accession number HGS_A135, but is not identical thereto.

[0642] 21. Full-length PRO4422 Polypeptides

[0643] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4422 (shown in FIG. 42 andSEQ ID NO:76) has certain amino acid sequence identity with lysozyme g.Accordingly, it is presently believed that PRO4422 disclosed in thepresent application is a newly identified member of the lysozyme familyand may have lysozyme activy.

[0644] 22. Full-length PRO4430 Polypeptides

[0645] Using WU-BLAST2 sequence alignment computer programs, it has beenfound that a full-length native sequence PRO4430 (shown in FIG. 44 andSEQ ID NO:78) has certain amino acid sequence identity with the proteinin GenBank accession number MMHC213L3_(—)9. Accordingly, it is presentlybelieved that PRO4430 disclosed in the present application is related tothe GenBank protein and may share at least one similar mechanism.

[0646] 23. Full-length PRO4499 Polypeptides

[0647] As far as is known, the DNA96889-2641 sequence encodes a novelfactor designated herein as PRO4499; using WU-BLAST2 sequence alignmentcomputer programs, limited sequence identities to known proteins wererevealed.

[0648] B. PRO Polypeptide Variants

[0649] In addition to the full-length native sequence PRO polypeptidesdescribed herein, it is contemplated that PRO variants can be prepared.PRO variants can be prepared by introducing appropriate nucleotidechanges into the PRO DNA, and/or by synthesis of the desired PROpolypeptide. Those skilled in the art will appreciate that amino acidchanges may alter post-translational processes of the PRO, such aschanging the number or position of glycosylation sites or altering themembrane anchoring characteristics.

[0650] Variations in the native full-length sequence PRO or in variousdomains of the PRO described herein, can be made, for example, using anyof the techniques and guidelines for conservative and non-conservativemutations set forth, for instance, in U.S. Pat. No. 5,364,934.Variations may be a substitution, deletion or insertion of one or morecodons encoding the PRO that results in a change in the amino acidsequence of the PRO as compared with the native sequence PRO. Optionallythe variation is by substitution of at least one amino acid with anyother amino acid in one or more of the domains of the PRO. Guidance indetermining which amino acid residue may be inserted, substituted ordeleted without adversely affecting the desired activity may be found bycomparing the sequence of the PRO with that of homologous known proteinmolecules and minimizing the number of amino acid sequence changes madein regions of high homology. Amino acid substitutions can be the resultof replacing one amino acid with another amino acid having similarstructural and/or chemical properties, such as the replacement of aleucine with a serine, i.e., conservative amino acid replacements.Insertions or deletions may optionally be in the range of about 1 to 5amino acids. The variation allowed may be determined by systematicallymaking insertions, deletions or substitutions of amino acids in thesequence and testing the resulting variants for activity exhibited bythe full-length or mature native sequence.

[0651] PRO polypeptide fragments are provided herein. Such fragments maybe truncated at the N-terminus or C-terminus, or may lack internalresidues, for example, when compared with a full length native protein.Certain fragments lack amino acid residues that are not essential for adesired biological activity of the PRO polypeptide.

[0652] PRO fragments may be prepared by any of a number of conventionaltechniques. Desired peptide fragments may be chemically synthesized. Analternative approach involves generating PRO fragments by enzymaticdigestion, e.g., by treating the protein with an enzyme known to cleaveproteins at sites defined by particular amino acid residues, or bydigesting the DNA with suitable restriction enzymes and isolating thedesired fragment. Yet another suitable technique involves isolating andamplifying a DNA fragment encoding a desired polypeptide fragment, bypolymerase chain reaction (PCR). Oligonucleotides that define thedesired termini of the DNA fragment are employed at the 5′ and 3′primers in the PCR. Preferably, PRO polypeptide fragments share at leastone biological and/or immunological activity with the native PROpolypeptide disclosed herein.

[0653] In particular embodiments, conservative substitutions of interestare shown in Table 6 under the heading of preferred substitutions. Ifsuch substitutions result in a change in biological activity, then moresubstantial changes, denominated exemplary substitutions in Table 6, oras further described below in reference to amino acid classes, areintroduced and the products screened. TABLE 6 Original ExemplaryPreferred Residue Substitutions Substitutions Ala (A) val; leu; ile valArg (R) lys; gln; asn lys Asn (N) gln; his; lys; arg gln Asp (D) glu gluCys (C) ser ser Gln (Q) asn asn Glu (E) asp asp Gly (G) pro; ala ala His(H) asn; gln; lys; arg arg Ile (I) leu; val; met; ala; phe; leunorleucine Leu (L) norleucine; ile; val; ile met; ala; phe Lys (K) arg;gln; asn arg Met (M) leu; phe; ile leu Phe (F) leu; val; ile; ala; tyrleu Pro (P) ala ala Ser (S) thr thr Thr (T) ser ser Trp (W) tyr; phe tyrTyr (Y) trp; phe; thr; ser phe Val (V) ile; leu; met; phe; leu ala;norleucine

[0654] Substantial modifications in function or immunological identityof the PRO polypeptide are accomplished by selecting substitutions thatdiffer significantly in their effect on maintaining (a) the structure ofthe polypeptide backbone in the area of the substitution, for example,as a sheet or helical conformation, (b) the charge or hydrophobicity ofthe molecule at the target site, or (c) the bulk of the side chain.Naturally occurring residues are divided into groups based on commonside-chain properties:

[0655] (1) hydrophobic: norleucine, met, ala, val, leu, ile;

[0656] (2) neutral hydrophilic: cys, ser, thr;

[0657] (3) acidic: asp, glu;

[0658] (4) basic: asn, gin, his, lys, arg;

[0659] (5) residues that influence chain orientation: gly, pro; and

[0660] (6) aromatic: trp, tyr, phe.

[0661] Non-conservative substitutions will entail exchanging a member ofone of these classes for another class. Such substituted residues alsomay be introduced into the conservative substitution sites or, morepreferably, into the remaining (non-conserved) sites.

[0662] The variations can be made using methods known in the art such asoligonucleotide-mediated (site-directed) mutagenesis, alanine scanning,and PCR mutagenesis. Site-directed mutagenesis [Carter et al., Nucl.Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res., 10:6487(1987)], cassette mutagenesis [Wells et al., Gene, 34:315 (1985)],restriction selection mutagenesis [Wells et al., Philos. Trans. R. Soc.London SerA, 317:415 (1986)] or other known techniques can be performedon the cloned DNA to produce the PRO variant DNA.

[0663] Scanning amino acid analysis can also be employed to identify oneor more amino acids along a contiguous sequence. Among the preferredscanning amino acids are relatively small, neutral amino acids. Suchamino acids include alanine, glycine, serine, and cysteine. Alanine istypically a preferred scanning amino acid among this group because iteliminates the side-chain beyond the beta-carbon and is less likely toalter the main-chain conformation of the variant [Cunningham and Wells,Science, 244: 1081-1085 (1989)]. Alanine is also typically preferredbecause it is the most common amino acid. Further, it is frequentlyfound in both buried and exposed positions [Creighton, The Proteins,(W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150:1 (1976)]. Ifalanine substitution does not yield adequate amounts of variant, anisoteric amino acid can be used.

[0664] C. Modifications of PRO

[0665] Covalent modifications of PRO are included within the scope ofthis invention. One type of covalent modification includes reactingtargeted amino acid residues of a PRO polypeptide with an organicderivatizing agent that is capable of reacting with selected side chainsor the N- or C-terminal residues of the PRO. Derivalization withbifunctional agents is useful, for instance, for crosslinking PRO to awater-insoluble support matrix or surface for use in the method forpurifying anti-PRO antibodies, and vice-versa. Commonly usedcrosslinking agents include, e.g., 1,1-bis(diazoacetyl)-2-phenylethane,glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with4-azidosalicylic acid, homobifunctional imidoesters, includingdisuccinimidyl esters such as 3,3′-dithiobis(succinimidylpropionate),bifunctional maleimides such as bis-N-maleimido-1,8-octane and agentssuch as methyl-3-[(p-azidophenyl)dithio]propioimidate.

[0666] Other modifications include deamidation of glutaminyl andasparaginyl residues to the corresponding glutamyl and aspartylresidues, respectively, hydroxylation of proline and lysine,phosphorylation of hydroxyl groups of seryl or threonyl residues,methylation of the α-amino groups of lysine, arginine, and histidineside chains [T. E. Creighton, Proteins: Structure and MolecularProperties, W.H. Freeman & Co., San Francisco, pp. 79-86 (1983)],acetylation of the N-terminal amine, and amidation of any C-terminalcarboxyl group.

[0667] Another type of covalent modification of the PRO polypeptideincluded within the scope of this invention comprises altering thenative glycosylation pattern of the polypeptide. “Altering the nativeglycosylation pattern” is intended for purposes herein to mean deletingone or more carbohydrate moieties found in native sequence PRO (eitherby removing the underlying glycosylation site or by deleting theglycosylation by chemical and/or enzymatic means), and/or adding one ormore glycosylation sites that are not present in the native sequencePRO. In addition, the phrase includes qualitative changes in theglycosylation of the native proteins, involving a change in the natureand proportions of the various carbohydrate moieties present.

[0668] Addition of glcosylation sites to the PRO polypeptide may beaccomplished by altering the amino acid sequence. The alteration may bemade, for example, by the addition of, or substitution by, one or moreserine or threonine residues to the native sequence PRO (for O-linkedglycosylation sites). The PRO amino acid sequence may optionally bealtered through changes at the DNA level, particularly by mutating theDNA encoding the PRO polypeptide at preselected bases such that codonsare generated that will translate into the desired amino acids.

[0669] Another means of increasing the number of carbohydrate moietieson the PRO polypeptide is by chemical or enzymatic coupling ofglycosides to the polypeptide. Such methods are described in the art,e.g., in WO 87/05330 published Sep. 11, 1987, and in Aplin and Wriston,CRC Crit. Rev. Biochem., pp. 259-306 (1981).

[0670] Removal of carbohydrate moieties present on the PRO polypeptidemay be accomplished chemically or enzymatically or by mutationalsubstitution of codons encoding for amino acid residues that serve astargets for glycosylation. Chemical deglycosylation techniques are knownin the art and described, for instance, by Hakimuddin, et al., Arch.Biochem. Biophys., 259:52 (1987) and by Edge et al., Anal. Biochem.,118:131 (1981). Enzymatic cleavage of carbohydrate moieties onpolypeptides can be achieved by the use of a variety of endo- andexo-glycosidases as described by Thotakura et al., Meth. Enzymol.,138:350 (1987).

[0671] Another type of covalent modification of PRO comprises linkingthe PRO polypeptide to one of a variety of nonproteinaceous polymers,e.g., polyethylene glycol (PEG), polypropylene glycol, orpolyoxyalkylenes, in the manner set forth in U.S. Pat. Nos. 4,640,835;4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

[0672] The PRO of the present invention may also be modified in a way toform a chimeric molecule comprising PRO fused to another, heterologouspolypeptide or amino acid sequence.

[0673] In one embodiment, such a chimeric molecule comprises a fusion ofthe PRO with a tag polypeptide which provides an epitope to which ananti-tag antibody can selectively bind. The epitope tag is generallyplaced at the amino- or carboxyl-terminus of the PRO. The presence ofsuch epitope-tagged forms of the PRO can be detected using an antibodyagainst the tag polypeptide. Also, provision of the epitope tag enablesthe PRO to be readily purified by affinity purification using ananti-tag antibody or another type of affinity matrix that binds to theepitope tag. Various tag polypeptides and their respective antibodiesare well known in the art. Examples include poly-histidine (poly-his) orpoly-histidine-glycine (poly-his-gly) tags; the flu HA tag polypeptideand its antibody 12CA5 [Field et al., Mol. Cell. Biol., 8:2159-2165(1988)]; the c-myc tag and the 8F9, 3C7, 6E10, G4, B7 and 9E10antibodies thereto [Evan et al., Molecular and Cellular Biology,5:3610-3616 (1985)]; and the Herpes Simplex virus glycoprotein D (gD)tag and its antibody [Paborsky et al., Protein Engineering, 3(6):547-553(1990)]. Other tag polypeptides include the Flag-peptide [Hopp et al.,BioTechnology, 6:1204-1210 (1988)]; the KT3 epitope peptide [Martin etal., Science, 255:192-194 (1992)]; an α-tubulin epitope peptide [Skinneret al., J. Biol. Chem., 266:15163-15166 (1991)]; and the T7 gene 10protein peptide tag [Lutz-Freyermuth et al., Proc. Natl. Acad. Sci. USA,87:6393-6397 (1990)].

[0674] In an alternative embodiment, the chimeric molecule may comprisea fusion of the PRO with an immunoglobulin or a particular region of animmunoglobulin. For a bivalent form of the chimeric molecule (alsoreferred to as an “immunoadhesin”), such a fusion could be to the Fcregion of an IgG molecule. The Ig fusions preferably include thesubstitution of a soluble (transmembrane domain deleted or inactivated)form of a PRO polypeptide in place of at least one variable regionwithin an Ig molecule. In a particularly preferred embodiment, theimmunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge,CH1, CH2 and CH3 regions of an IgG1 molecule. For the production ofimmunoglobulin fusions see also U.S. Pat. No. 5,428,130 issued Jun. 27,1995.

[0675] D. Preparation of PRO

[0676] The description below relates primarily to production of PRO byculturing cells transformed or transfected with a vector containing PROnucleic acid. It is, of course, contemplated that alternative methods,which are well known in the art, may be employed to prepare PRO. Forinstance, the PRO sequence, or portions thereof, may be produced bydirect peptide synthesis using solid-phase techniques [see, e.g.,Stewart et al., Solid-Phase Peptide Synthesis, W.H. Freeman Co., SanFrancisco, Calif. (1969); Merrifield, J. Am. Chem. Soc., 85:2149-2154(1963)]. In vitro protein synthesis may be performed using manualtechniques or by automation. Automated synthesis may be accomplished,for instance, using an Applied Biosystems Peptide Synthesizer (FosterCity, Calif.) using manufacturer's instructions. Various portions of thePRO may be chemically synthesized separately and combined using chemicalor enzymatic methods to produce the full-length PRO.

[0677] 1. Isolation of DNA Encoding PRO

[0678] DNA encoding PRO may be obtained from a cDNA library preparedfrom tissue believed to possess the PRO mRNA and to express it at adetectable level. Accordingly, human PRO DNA can be convenientlyobtained from a cDNA library prepared from human tissue, such asdescribed in the Examples. The PRO-encoding gene may also be obtainedfrom a genomic library or by known synthetic procedures (e.g., automatednucleic acid synthesis).

[0679] Libraries can be screened with probes (such as antibodies to thePRO or oligonucleotides of at least about 20-80 bases) designed toidentify the gene of interest or the protein encoded by it. Screeningthe cDNA or genomic library with the selected probe may be conductedusing standard procedures, such as described in Sambrook et al.,Molecular Cloning: A Laboratory Manual (New York: Cold Spring HarborLaboratory Press, 1989). An alternative means to isolate the geneencoding PRO is to use PCR methodology [Sambrook et al., supra;Dieffenbach et al., PCR Primer: A Laboratory Manual (Cold Spring HarborLaboratory Press, 1995)].

[0680] The Examples below describe techniques for screening a cDNAlibrary. The oligonucleotide sequences selected as probes should be ofsufficient length and sufficiently unambiguous that false positives areminimized. The oligonucleotide is preferably labeled such that it can bedetected upon hybridization to DNA in the library being screened.Methods of labeling are well known in the art, and include the use ofradiolabels like ³²P-labeled ATP, biotinylation or enzyme labeling.Hybridization conditions, including moderate stringency and highstringency, are provided in Sambrook et al., supra.

[0681] Sequences identified in such library screening methods can becompared and aligned to other known sequences deposited and available inpublic databases such as GenBank or other private sequence databases.Sequence identity (at either the amino acid or nucleotide level) withindefined regions of the molecule or across the full-length sequence canbe determined using methods known in the art and as described herein.

[0682] Nucleic acid having protein coding sequence may be obtained byscreening selected cDNA or genomic libraries using the deduced aminoacid sequence disclosed herein for the first time, and, if necessary,using conventional primer extension procedures as described in Sambrooket al., supra, to detect precursors and processing intermediates of mRNAthat may not have been reverse-transcribed into cDNA.

[0683] 2. Selection and Transformation of Host Cells

[0684] Host cells are transfected or transformed with expression orcloning vectors described herein for PRO production and cultural inconventional nutrient media modified as appropriate for inducingpromoters, selecting transformants, or amplifying the genes encoding thedesired sequences. The culture conditions, such as media, temperature,pH and the like, can be selected by the skilled artisan without undueexperimentation. In general, principles, protocols, and practicaltechniques for maximizing the productivity of cell cultures can be foundin Mammalian Cell Biotechnology: a Practical Approach, M. Butler, ed.(IRL Press, 1991) and Sambrook et al., supra.

[0685] Methods of eulkaryotic cell transfection and prokaryotic celltransformation are known to the ordinarily skilled artisan, for example,CaCl₂, CaPO₄, liposome-mediated and electroporation. Depending on thehost cell used, transformation is performed using standard techniquesappropriate to such cells. The calcium treatment employing calciumchloride, as described in Sambrook et al., supra, or electroporation isgenerally used for prokaryotes. Infection with Agrobacterium tumefaciensis used for transformation of certain plant cells, as described by Shawet al., Gene, 23:315 (1983) and WO 89/05859 published Jun. 29, 1989. Formammalian cells without such cell walls, the calcium phosphateprecipitation method of Graham and van der Eb, Virology, 52:456457(1978) can be employed. General aspects of mammalian cell host systemtransfections have been described in U.S. Pat. No. 4,399,216.Transformations into yeast are typically carried out according to themethod of Van Solingen et al., J. Bact., 130:946 (1977) and Hsiao etal., Proc. Natl. Acad. Sci. (USA), 76:3829 (1979). However, othermethods for introducing DNA into cells, such as by nuclearmicroinjection, electroporation, bacterial protoplast fusion with intactcells, or polycations, e.g., polybrene, polyornithine, may also be used.For various techniques for transforming mammalian cells, see Keown etal., Methods in Enzymology, 185:527-537 (1990) and Mansour et al.,Nature, 336:348-352 (1988).

[0686] Suitable host cells for cloning or expressing the DNA in thevectors herein include prokaryote, yeast, or higher eukaryote cells.Suitable prokaryotes include but are not limited to eubacteria, such asGram-negative or Gram-positive organisms, for example,Enterobacteriaceae such as E. coli. Various E. coli strains are publiclyavailable, such as E. coli K12 strain MM294 (ATCC 31,446); E. coli X1776(ATCC 31,537); E. coli strain W3110 (ATCC 27,325) and K5772 (ATCC53,635). Other suitable prokaryotic host cells includeEnterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter,Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium,Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacillisuch as B. subtilis and B. licheniformis (e.g., B. licheniformis 41Pdisclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P.aeruginosa, and Streptonyces. These examples are illustrative ratherthan limiting. Strain W3110 is one particularly preferred host or parenthost because it is a common host strain for recombinant DNA productfermentations. Preferably, the host cell secretes minimal amounts ofproteolytic enzymes. For example, strain W3110 may be modified to effecta genetic mutation in the genes encoding proteins endogenous to thehost, with examples of such hosts including E. coli W3110 strain 1A2,which has the complete genotype tonA; E. coli W3110 strain 9E4, whichhas the complete genotype tonA ptr3; E. coli W3110 strain 27C7 (ATCC55,244), which has the complete genotype tonA ptr3phoA E15 (argF-lac)169degP ompTkan^(r) ; E. coli W3110 strain 37D6, which has the completegenotype tonA ptr3 phoA E15 (argF-lac)169 degP ompT rbs7 ilvG kan^(r) ;E. coli W3110 strain 40B4, which is strain 37D6 with a non-kanamycinresistant degP deletion mutation; and an E. coli strain having mutantperiplasmic protease disclosed in U.S. Pat. No. 4,946,783 issued Aug. 7,1990. Alternatively, in vitro methods of cloning, e.g., PCR or othernucleic acid polymerase reactions, are suitable.

[0687] In addition to prokaryotes, eukaryotic microbes such asfilamentous fungi or yeast are suitable cloning or expression hosts forPRO encoding vectors. Saccharonyces cerevisiae is a commonly used lowereukaryotic host microorganism. Others include Schizosaccharomyces pombe(Beach and Nurse, Nature, 290: 140 [1981]; EP 139,383 published May 2,1985); Kluyveronyces hosts (U.S. Pat. No. 4,943,529; Fleer et al.,Bio/Technology, 9:968-975 (1991)) such as, e.g., K. lactis (MW98-8C,CBS683, CBS4574; Louvencourt et al., J. Bacteriol., 154(2):737-742[1983]), K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K.wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum(ATCC 36,906; Van den Berg et al., Bio/Technology, 8:135 (1990)), K.thernotolerans, and K. marxianus; yarrowia (EP 402,226); Pichiapastoris(EP 183,070; Sreekrislna et al., J. Basic Microbiol., 28:265-278[1988]); Candida; Trichodenna reesia (EP 244,234); Neurospora crassa(Case et al., Proc. Natl. Acad. Sci. USA, 76:5259-5263 [1979]);Schwanniomyces such as Schwanniomyces occidentalis (EP 394,538 publishedOct. 31, 1990); and filamentous fungi such as, e.g., Neurospora,Penicillium, Tolypocladium (WO 91/00357 published Jan. 10, 1991), andAspergillus hosts such as A. nidulans (Ballance et al., Biochem.Biophys. Res. Commun., 112:284-289 [1983]; Tilbum et al., Gene,26:205-221 [1983]; Yelton et al., Proc. Natl. Acad. Sci. USA, 81:1470-1474 [1984]) and A. niger (Kelly and Hynes, EMBO J., 4:475-479[1985]). Methylotropic yeasts are suitable herein and include, but arenot limited to, yeast capable of growth on methanol selected from thegenera consisting of Hansenula, Candida, Kloeckera, Pichia,Saccharomyces, Torulopsis, and Rhodotorula. A list of specific speciesthat are exemplary of this class of yeasts may be found in C. Anthony,The Biochemistry of Methylotrophs, 269 (1982).

[0688] Suitable host cells for the expression of glycosylated PRO arederived from multicellular organisms. Examples of invertebrate cellsinclude insect cells such as Drosophila S2 and Spodoptera Sf9, as wellas plant cells. Examples of useful mammalian host cell lines includeChinese hamster ovary (CHO) and COS cells. More specific examplesinclude monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL1651); human embryonic kidney line (293 or 293 cells subcloned forgrowth in suspension culture, Graham et al., J. Gen Virol., 36:59(1977)); Chinese hamster ovary cells/-DHFR (CHO, Urlaub and Chasin,Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells (TM4,Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138, ATCCCCL 75); human liver cells (Hep G2,HB 8065); and mouse mammary tumor(MMT 060562, ATCC CCL51). The selection of the appropriate host cell isdeemed to be within the skill in the art.

[0689] 3. Selection and Use of a Replicable Vector

[0690] The nucleic acid (e.g., cDNA or genomic DNA) encoding PRO may beinserted into a replicable vector for cloning (amplification of the DNA)or for expression. Various vectors are publicly available. The vectormay, for example, be in the form of a plasmid, cosmid, viral particle,or phage. The appropriate nucleic acid sequence may be inserted into thevector by a variety of procedures. In general, DNA is inserted into anappropriate restriction endonuclease site(s) using techniques known inthe art. Vector components generally include, but are not limited to,one or more of a signal sequence, an origin of replication, one or moremarker genes, an enhancer element, a promoter, and a transcriptiontermination sequence. Construction of suitable vectors containing one ormore of these components employs standard ligation techniques which areknown to the skilled artisan.

[0691] The PRO may be produced recombinantly not only directly, but alsoas a fusion polypeptide with a heterologous polypeptilde, which may be asignal sequence or other polypeptide having a specific cleavage site atthe N-terminus of the mature protein or polypeptide. In general, thesignal sequence may be a component of the vector, or it may be a part ofthe PRO-encoding DNA that is inserted into the vector. The signalsequence may be a prokaryotic signal sequence selected, for example,from the group of the alkaline phosphatase, penicillinase, lpp, orheat-stable enterotoxin II leaders. For yeast secretion the signalsequence may be, e.g., the yeast invertase leader, alpha factor leader(including Saccharomyces and Kluyveromyces α-factor leaders, the latterdescribed in U.S. Pat. No. 5,010,182), or acid phosphatase leader, theC. albicans glucoamylase leader (EP 362,179 published Apr. 4, 1990), orthe signal described in WO 90/13646 published Nov. 15, 1990. Inmammalian cell expression, mammalian signal sequences may be used todirect secretion of the protein, such as signal sequences from secretedpolypeptides of the same or related species, as well as viral secretoryleaders.

[0692] Both expression and cloning vectors contain a nucleic acidsequence that enables the vector to replicate in one or more selectedhost cells. Such sequences are well known for a variety of bacteria,yeast, and viruses. The origin of replication from the plasmid pBR322 issuitable for most Gram-negative bacteria, the 2μ plasmid origin issuitable for yeast, and various viral origins (SV40, polyoma,adenovirus, VSV or BPV) are useful for cloning vectors in mammaliancells.

[0693] Expression and cloning vectors will typically contain a selectiongene, also termed a selectable marker. Typical selection genes encodeproteins that (a) confer resistance to antibiotics or other toxins,e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b)complement auxotrophic deficiencies, or (c) supply critical nutrientsnot available from complex media, e.g., the gene encoding D-alanineracemase for Bacilli.

[0694] An example of suitable selectable markers for mammalian cells arethose that enable the identification of cells competent to take up thePRO-encoding nucleic acid, such as DHFR or thymidine kinase. Anappropriate host cell when wild-type DHFR is employed is the CHO cellline deficient in DHFR activity, prepared and propagated as described byUrlaub etal., Proc. Natl. Acad. Sci. USA, 77:4216(1980). A suitableselection gene for use in yeast is the trpl gene present in the yeastplasmid YRp7 [Stinchcomb et al., Nature, 282:39 (1979); Kingsman et al.,Gene, 7:141 (1979); Tschemper et al., Gene, 10: 157 (1980)]. The trp1gene provides a selection marker for a mutant strain of yeast lackingthe ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1[Jones, Genetics, 85:12 (1977)].

[0695] Expression and cloning vectors usually contain a promoteroperably linked to the PRO-encoding nucleic acid sequence to direct mRNAsynthesis. Promoters recognized by a variety of potential host cells arewell known. Promoters suitable for use with prokaryotic hosts includethe β-lactamase and lactose promoter systems [Chang et al., Nature,275:615 (1978); Goeddel et al., Nature, 281:544 (1979)], alkalinephosphatase, a tryptophan (trp) promoter system [Goeddel, Nucleic AcidsRes., 8:4057 (1980); EP 36,776], and hybrid promoters such as the tacpromoter [deBoer et al., Proc. Natl. Acad. Sci. USA, 80:21-25 (1983)].Promoters for use in bacterial systems also will contain a Shine-Dalgamo(S.D.) sequence operably linked to the DNA encoding PRO.

[0696] Examples of suitable promoting sequences for use with yeast hostsinclude the promoters for 3-phosphoglycerate kinase [Hitzeman et al., J.Biol. Chem., 255:2073 (1980)] or other glycolytic enzymes [Hess et al.,J. Adv. Enzyme Reg., 7:149 (1968); Holland, Biochemistry, 17:4900(1978)], such as enolase, glyceraldehyde-3-phosphate dehydrogenase,hexokinase, pyruvate decarboxylase, phosphofructokinase,glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvatekinase, triosephosphate isomerase, phosphoglucose isomerase, andglucokinase.

[0697] Other yeast promoters, which are inducible promoters having theadditional advantage of transcription controlled by growth conditions,are the promoter regions for alcohol dehydrogenase 2, isocytochrome C,acid phosphatase, degradative enzymes associated with nitrogenmetabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase,and enzymes responsible for maltose and galactose utilization. Suitablevectors and promoters for use in yeast expression are further describedin EP 73,657.

[0698] PRO transcription from vectors in mammalian host cells iscontrolled, for example, by promoters obtained from the genomes ofviruses such as polyoma virus, fowlpox virus (UK 2,211,504 publishedJul. 5, 1989), adenovirus (such as Adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-Bvirus and Simian Virus 40 (SV40), from heterologous mammalian promoters,e.g., the actin promoter or an iramunoglobulin promoter, and fromheat-shock promoters, provided such promoters are compatible with thehost cell systems.

[0699] Transcription of a DNA encoding the PRO by higher eukaryotes maybe increased by inserting an enhancer sequence into the vector.Enhancers are cis-acting elements of DNA, usually about from 10 to 300bp, that act on a promoter to increase its transcription. Many enhancersequences are now known from mammalian genes (glolbin, elastase,albumin, a-fetoprotein, and insulin). Typically, however, one will usean enhancer from a eukaryotic cell virus. Examples include the SV40enhancer on the late side of the replication origin (bp 100-270), thecytomegalovirus early promoter enhancer, the polyoma enhancer on thelate side of the replication origin, and adenovirus enhancers. Theenhancer may be spliced into the vector at a position 5′ or 3′ to thePRO coding sequence, but is preferably located at a site 5′ from thepromoter.

[0700] Expression vectors used in eukaryotic host cells (yeast, fingi,insect, plant, animal, human, or nucleated cells from othermulticellular organisms) will also contain sequences necessary for thetermination of transcription and for stabilizing the mRNA. Suchsequences are commonly available from the 5′ and, occasionally 3′,untranslated regions of eukaryotic or viral DNAs or cDNAs. These regionscontain nucleotide segments transcribed as polyadenylated fragments inthe untranslated portion of the mRNA encoding PRO.

[0701] Still other methods, vectors, and host cells suitable foradaptation to the synthesis of PRO in recombinant vertebrate cellculture are described in Gething et al., Nature, 293:620-625 (1981);Mantei et al., Nature, 281:40-46 (1979); EP 117,060; and EP 117,058.

[0702] 4. Detecting Gene Amplification/Expression

[0703] Gene amplification and/or expression may be measured in a sampledirectly, for example, by conventional Southern blotting, Northernblotting to quantitate the transcription of mRNA [Thomas, Proc. Natl.Acad. Sci. USA, 77:5201-5205 (1980)], dot blotting (DNA analysis), or insitu hybridization, using an appropriately labeled probe, based on thesequences provided herein. Alternatively, antibodies may be employedthat can recognize specific duplexes, including DNA duplexes, RNAduplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. Theantibodies in turn may be labeled and the assay may be carried out wherethe duplex is bound to a surface, so that upon the formation of duplexon the surface, the presence of antibody bound to the duplex can bedetected.

[0704] Gene expression, alternatively, may be measured by immunologicalmethods, such as immunohistochemical staining of cells or tissuesections and assay of cell culture or body fluids, to quantitatedirectly the expression of gene product. Antibodies useful forimmunohistochemical staining and/or assay of sample fluids may be eithermonoclonal or polyclonal, and may be prepared in any mammal.Conveniently, the antibodies may be prepared against a native sequencePRO polypeptide or against a synthetic peptide based on the DNAsequences provided herein or against exogenous sequence fused to PRO DNAand encoding a specific antibody epitope.

[0705] 5. Purification of Polypeptide

[0706] Forms of PRO may be recovered from culture medium or from hostcell lysates. If membrane-bound, it can be released from the membraneusing a suitable detergent solution (e.g. Triton-X 100) or by enzymaticcleavage. Cells employed in expression of PRO can be disrupted byvarious physical or chemical means, such as freeze-thaw cycling,sonication, mechanical disruption, or cell lysing agents.

[0707] It may be desired to purify PRO from recombinant cell proteins orpolypeptides. The following procedures are exemplary of suitablepurification procedures: by fractionation on an ion-exchange column;ethanol precipitation; reverse phase HPLC; chromatography on silica oron a cation-exchange resin such as DEAE; chromatofocusing; SDS-PAGE;ammonium sulfate precipitation; gel filtration using, for example,Sephadex G-75; protein A Sepharose columns to remove contaminants suchas IgG; and metal chelating columns to bind epitope-tagged forms of thePRO. Various methods of protein purification may be employed and suchmethods are known in the art and described for example in Deutscher,Methods in Enzymology, 182 (1990); Scopes, Protein Purification:Principles and Practice, Springer-Verlag, New York (1982). Thepurification step(s) selected will depend, for example, on the nature ofthe production process used and the particular PRO produced.

[0708] E. Uses for PRO

[0709] Nucleotide sequences (or their complement) encoding PRO havevarious applications in the art of molecular biology, including uses ashybridization probes, in chromosome and gene mapping and in thegeneration of anti-sense RNA and DNA. PRO nucleic acid will also beuseful for the preparation of PRO polypeptides by the recombinanttechniques described herein.

[0710] The full-length native sequence PRO gene, or portions thereof,may be used as hybridization probes for a cDNA library to isolate thefull-length PRO cDNA or to isolate still other cDNAs (for instance,those encoding naturally-occurring variants of PRO or PRO from otherspecies) which have a desired sequence identity to the native PROsequence disclosed herein. Optionally, the length of the probes will beabout 20 to about 50 bases. The hybridization probes may be derived fromat least partially novel regions of the full length native nucleotidesequence wherein those regions may be determined without undueexperimentation or from genomic sequences including promoters, enhancerelements and introns of native sequence PRO. By way of example, ascreening method will comprise isolating the coding region of the PROgene using the known DNA sequence to synthesize a selected probe ofabout 40 bases. Hybridization probes may be labeled by a variety oflabels, including radionucleotides such as ³²p or ³⁵S, or enzymaticlabels such as alkaline phosphatase coupled to the probe viaavidin/biotin coupling systems. Labeled probes having a sequencecomplementary to that of the PRO gene of the present invention can beused to screen libraries of human cDNA, genomic DNA or mRNA to determinewhich members of such libraries the probe hybridizes to. Hybridizationtechniques are described in further detail in the Examples below.

[0711] Any EST sequences disclosed in the present application maysimilarly be employed as probes, using the methods disclosed herein.

[0712] Other useful fragments of the PRO nucleic acids include antisenseor sense oligonucleotides comprising a singe-stranded nucleic acidsequence (either RNA or DNA) capable of binding to target PRO mRNA(sense) or PRO DNA (antisense) sequences. Antisense or senseoligonucleotides, according to the present invention, comprise afragment of the coding region of PRO DNA. Such a fragment generallycomprises at least about 14 nucleotides, preferably from about 14 to 30nucleotides. The ability to derive an antisense or a senseoligonucleotide, based upon a cDNA sequence encoding a given protein isdescribed in, for example, Stein and Cohen (Cancer Res. 48:2659, 1988)and van der Krol et al. (BioTechniques 6:958, 1988).

[0713] Binding of antisense or sense oligonucleotides to target nucleicacid sequences results in the formation of duplexes that blocktranscription or translation of the target sequence by one of severalmeans, including enhanced degradation of the duplexes, prematuretermination of transcription or translation, or by other means. Theantisense oligonucleotides thus may be used to block expression of PROproteins. Antisense or sense oligonucleotides further compriseoligonucleotides having modified sugar-phosphodiester backbones (orother sugar linkages, such as those described in WO 91/06629) andwherein such sugar linkages are resistant to endogenous nucleases. Sucholigonucleotides with resistant sugar linkages are stable in vivo (i.e.,capable of resisting enzymatic degradation) but retain sequencespecificity to be able to bind to target nucleotide sequences.

[0714] Other examples of sense or antisense oligonucleotides includethose oligonucleotides which are covalently linked to organic moieties,such as those described in WO 90/10048, and other moieties thatincreases affinity of the oligonucleotide for a target nucleic acidsequence, such as poly-(L-lysine). Further still, intercalating agents,such as ellipticine, and alkylating agents or metal complexes may beattached to sense or antisense oligonucleotides to modify bindingspecificities of the antisense or sense oligonucleotide for the targetnucleotide sequence.

[0715] Antisense or sense oligonucleotides may be introduced into a cellcontaining the target nucleic acid sequence by any gene transfer method,including, for example, CaPO₄-mediated DNA transfection,electroporation, or by using gene transfer vectors such as Epstein-Barrvirus. In a preferred procedure, an antisense or sense oligonucleotideis inserted into a suitable retroviral vector. A cell containing thetarget nucleic acid sequence is contacted with the recombinantretroviral vector, either in vivo or ex vivo. Suitable retroviralvectors include, but are not limited to, those derived from the murineretrovirus M-MuLV, N2 (a retrovirus derived from M-MuLV), or the doublecopy vectors designated DCT5A, DCT5B and DCT5C (see WO 90/13641).

[0716] Sense or antisense oligonucleotides also may be introduced into acell containing the target nucleotide sequence by formation of aconjugate with a ligand binding molecule, as described in WO 91/04753.Suitable ligand binding molecules include, but are not limited to, cellsurface receptors, growth factors, other cytokines, or other ligandsthat bind to cell surface receptors. Preferably, conjugation of theligand binding molecule does not substantially interfere with theability of the ligand binding molecule to bind to its correspondingmolecule or receptor, or block entry of the sense or antisenseoligonucleotide or its conjugated version into the cell.

[0717] Alternatively, a sense or an antisense oligonucleotide may beintroduced into a cell containing the target nucleic acid sequence byformation of an oligonucleotide-lipid complex, as described in WO90/10448. The sense or antisense oligonucleotide-lipid complex ispreferably dissociated within the cell by an endogenous lipase.

[0718] Antisense or sense RNA or DNA molecules are generally at leastabout 5 bases in length, about 10 bases in length, about 15 bases inlength, about 20 bases in length, about 25 bases in length, about 30bases in length, about 35 bases in length, about 40 bases in length,about 45 bases in length, about 50 bases in length, about 55 bases inlength, about 60 bases in length, about 65 bases in length, about 70bases in length, about 75 bases in length, about 80 bases in length,about 85 bases in length, about 90 bases in length, about 95 bases inlength, about 100 bases in length, or more.

[0719] The probes may also be employed in PCR techniques to generate apool of sequences for identification of closely related PRO codingsequences.

[0720] Nucleotide sequences encoding a PRO can also be used to constructhybridization probes for mapping the gene which encodes that PRO and forthe genetic analysis of individuals with genetic disorders. Thenucleotide sequences provided herein may be mapped to a chromosome andspecific regions of a chromosome using known techniques, such as in situhybridization, linkage analysis against known chromosomal markers, andhybridization screening with libraries.

[0721] When the coding sequences for PRO encode a protein which binds toanother protein (example, where the PRO is a receptor), the PRO can beused in assays to identify the other proteins or molecules involved inthe binding interaction. By such methods, inhibitors of thereceptor/ligand binding interaction can be identified. Proteins involvedin such binding interactions can also be used to screen for peptide orsmall molecule inhibitors or agonists of the binding interaction. Also,the receptor PRO can be used to isolate correlative ligand(s). Screeningassays can be designed to find lead compounds that mimic the biologicalactivity of a native PRO or a receptor for PRO. Such screening assayswill include assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates. Small molecules contemplated include syntheticorganic or inorganic compounds. The assays can be performed in a varietyof formats, including protein-protein binding assays, biochemicalscreening assays, immunoassays and cell based assays, which are wellcharacterized in the art.

[0722] Nucleic acids which encode PRO or its modified forms can also beused to generate either transgenic animals or “knock out” animals which,in turn, are useful in the development and screening of therapeuticallyuseful reagents. A transgenic animal (e.g., a mouse or rat) is an animalhaving cells that contain a transgene, which transgene was introducedinto the animal or an ancestor of the animal at a prenatal, e.g., anembryonic stage. A transgene is a DNA which is integrated into thegenome of a cell from which a transgenic animal develops. In oneembodiment, cDNA encoding PRO can be used to clone genomic DNA encodingPRO in accordance with established techniques and the genomic sequencesused to generate transgenic animals that contain cells which express DNAencoding PRO. Methods for generating transgenic animals, particularlyanimals such as mice or rats, have become conventional in the art andare described, for example, in U.S. Pat. Nos. 4,736,866 and 4,870,009.Typically, particular cells would be targeted for PRO transgeneincorporation with tissue-specific enhancers. Transgenic animals thatinclude a copy of a transgene encoding PRO introduced into the germ lineof the animal at an embryonic stage can be used to examine the effect ofincreased expression of DNA encoding PRO. Such animals can be used astester animals for reagents thought to confer protection from, forexample, pathological conditions associated with its overexpression. Inaccordance with this facet of the invention, an animal is treated withthe reagent and a reduced incidence of the pathological condition,compared to untreated animals bearing the transgene, would indicate apotential therapeutic intervention for the pathological condition.

[0723] Alternatively, non-human homologues of PRO can be used toconstruct a PRO “knock out” animal which has a defective or altered geneencoding PRO as a result of homologous recombination between theendogenous gene encoding PRO and altered genomic DNA encoding PROintroduced into an embryonic stem cell of the animal. For example, cDNAencoding PRO can be used to clone genomic DNA encoding PRO in accordancewith established techniques. A portion of the genomic DNA encoding PROcan be deleted or replaced with another gene, such as a gene encoding aselectable marker which can be used to monitor integration. Typically,several kilobases of unaltered flanking DNA (both at the 5′ and 3′ ends)are included in the vector [see e.g., Thomas and Capecchi, Cell, 51:503(1987) for a description of homologous recombination vectors]. Thevector is introduced into an embryonic stem cell line (e.g., byelectroporation) and cells in which the introduced DNA has homologouslyrecombined with the endogenous DNA are selected [see e.g., Li et al.,Cell, 69:915 (1992)]. The selected cells are then injected into ablastocyst of an animal (e.g., a mouse or rat) to form aggregationchimeras [see e.g., Bradley, in Teratocarcinomas and Embryonic StemCells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987),pp. 113-152]. A chimeric embryo can then be implanted into a suitablepseudopregnant female foster animal and the embryo brought to term tocreate a “knock out” animal. Progeny harboring the homologouslyrecombined DNA in their germ cells can be identified by standardtechniques and used to breed animals in which all cells of the animalcontain the homologously recombined DNA. Knockout animals can becharacterized for instance, for their ability to defend against certainpathological conditions and for their development of pathologicalconditions due to absence of the PRO polypeptide.

[0724] Nucleic acid encoding the PRO polypeptides may also be used ingene therapy. In gene therapy applications, genes are introduced intocells in order to achieve in vivo synthesis of a therapeuticallyeffective genetic product, for example for replacement of a defectivegene. “Gene therapy” includes both conventional gene therapy where alasting effect is achieved by a single treatment, and the administrationof gene therapeutic agents, which involves the one time or repeatedadministration of a therapeutically effective DNA or mRNA. AntisenseRNAs and DNAs can be used as therapeutic agents for blocking theexpression of certain genes in vivo. It has already been shown thatshort antisense oligonucleotides can be imported into cells where theyact as inhibitors, despite their low intracellular concentrations causedby their restricted uptake by the cell membrane. (Zamecnik et al., Proc.Natl. Acad. Sci. USA 83:4143-4146 [1986]). The oligonucleotides can bemodified to enhance their uptake, e.g. by substituting their negativelycharged phosphodiester groups by uncharged groups.

[0725] There are a variety of techniques available for introducingnucleic acids into viable cells. The techniques vary depending uponwhether the nucleic acid is transferred into cultured cells in vitro, orin vivo in the cells of the intended host. Techniques suitable for thetransfer of nucleic acid into mammalian cells in vitro include the useof liposomes, electroporation, microinjection, cell fusion,DEAE-dextran, the calcium phosphate precipitation method, erc. Thecurrently preferred in vivo gene transfer techniques includetransfection with viral (typically retroviral) vectors and viral coatprotein-liposome mediated transfection (Dzau et al., Trends inBiotechnology 11, 205-210 [1993]). In some situations it is desirable toprovide the nucleic acid source with an agent that targets the targetcells, such as an antibody specific for a cell surface membrane proteinor the target cell, a ligand for a receptor on the target cell, etc.Where liposomes are employed, proteins which bind to a cell surfacemembrane protein associated with endocytosis may be used for targetingand/or to facilitate uptake, e.g. capsid proteins or fragments thereoftropic for a particular cell type, antibodies for proteins which undergointernalization in cycling, proteins that target intracellularlocalization and enhance intracellular half-life. The technique ofreceptor-mediated endocytosis is described, for example, by Wu et al.,J. Biol. Chem. 262, 4429-4432 (1987); and Wagner et al., Proc. Natl.Acad. Sci. USA 87, 3410-3414 (1990). For review of gene marking and genetherapy protocols see Anderson et al., Science 256, 808-813 (1992).

[0726] The PRO polypeptides described herein may also be employed asmolecular weight markers for protein electrophoresis purposes and theisolated nucleic acid sequences may be used for recombinandy expressingthose markers.

[0727] The nucleic acid molecules encoding the PRO polypeptides orfragments thereof described herein are useful for chromosomeidentification. In this regard, there exists an ongoing need to identifynew chromosome markers, since relatively few chromosome markingreagents, based upon actual sequence data are presently available. EachPRO nucleic acid molecule of the present invention can be used as achromosome marker.

[0728] The PRO polypeptides and nucleic acid molecules of the presentinvention may also be used for tissue typing, wherein the PROpolypeptides of the present invention may be differentially expressed inone tissue as compared to another. PRO nucleic acid molecules will finduse for generating probes for PCR, Northern analysis, Southern analysisand Western analysis.

[0729] The PRO polypeptides described herein may also be employed astherapeutic agents. The PRO polypeptides of the present invention can beformulated according to known methods to prepare pharmaceutically usefulcompositions, whereby the PRO product hereof is combined in admixturewith a pharmaceutically acceptable carrier vehicle. Therapeuticformulations are prepared for storage by mixing the active ingredienthaving the desired degree of purity with optional physiologicallyacceptable carriers, excipients or stabilizers (Remington'sPharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the formof lyophilized formulations or aqueous solutions. Acceptable carriers,excipients or stabilizers are nontoxic to recipients at the dosages andconcentrations employed, and include buffers such as phosphate, citrateand other organic acids; antioxidants including ascorbic acid; lowmolecular weight (less than about 10 residues) polypeptides; proteins,such as serum albumin, gelatin or immunoglobulins; hydrophilic polymerssuch as polyvinylpyrrolidone, amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides and othercarbohydrates including glucose, mannose, or dextrins; chelating agentssuch as EDTA; sugar alcohols such as mannitol or sorbitol; salt-formingcounterions such as sodium; and/or nonionic surfactants such as TWEEN™,PLURONICS™ or PEG.

[0730] The formulations to be used for in vivo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes, prior to or following lyophilization andreconstitution.

[0731] Therapeutic compositions herein generally are placed into acontainer having a sterile access port, for example, an intravenoussolution bag or vial having a stopper pierceable by a hypodermicinjection needle.

[0732] The route of administration is in accord with known methods, e.g.injection or infusion by intravenous, intraperitoneal, intracerebral,intramuscular, intraocular, intraarterial or intralesional routes,topical administration, or by sustained release systems.

[0733] Dosages and desired drug concentrations of pharmaceuticalcompositions of the present invention may vary depending on theparticular use envisioned. The determination of the appropriate dosageor route of administration is well within the skill of an ordinaryphysician. Animal experiments provide reliable guidance for thedetermination of effective doses for human therapy. Interspecies scalingof effective doses can be performed following the principles laid downby Mordenti, J. and Chappell, W. “The use of interspecies scaling intoxicokinetics” In Toxicokinetics and New Drug Development, Yacobi etal., Eds., Pergamon Press, New York 1989, pp. 42-96.

[0734] When in vivo administration of a PRO polypeptide or agonist orantagonist thereof is employed, normal dosage amounts may vary fromabout 10 ng/kg to up to 100 mg/kg of mammal body weight or more per day,preferably about 1 μg/kg/day to 10 mg/kg/day, depending upon the routeof administration. Guidance as to particular dosages and methods ofdelivery is provided in the literature; see, for example, U.S. Pat. Nos.4,657,760; 5,206,344; or 5,225,212. It is anticipated that differentformulations will be effective for different treatment compounds anddifferent disorders, that administration targeting one organ or tissue,for example, may necessitate delivery in a manner different from that toanother organ or tissue.

[0735] Where sustained-release administration of a PRO polypeptide isdesired in a formulation with release characteristics suitable for thetreatment of any disease or disorder requiring administration of the PROpolypeptide, microencapsulation of the PRO polypeptide is contemplated.Microencapsulation of recombinant proteins for sustained release hasbeen successfully performed with human growth hormone (rhGH),interferon-(rhIFN-), interleukin-2, and MN rgp120. Johnson et al., Nat.Med., 2:795-799 (1996); Yasuda, Biomed. Ther., 27:1221-1223 (1993); Horaet al., Bio/Technology. 8:755-758 (1990); Cleland, “Design andProduction of Single Immunization Vaccines Using PolylactidePolyglycolide Microsphere Systems,” in Vaccine Design: The Subunit andAdjuvant Approach, Powell and Newman, eds, (Plenum Press: New York,1995), pp. 439-462; WO 97/03692, WO 96/40072, WO 96/07399; and U.S. Pat.No. 5,654,010.

[0736] The sustained-release formulations of these proteins weredeveloped using poly-lactic-coglycolic acid (PLGA) polymer due to itsbiocompatibility and wide range of biodegradable properties. Thedegradation products of PLGA, lactic and glycolic acids, can be clearedquickly within the human body. Moreover, the degradability of thispolymer can be adjusted from months to years depending on its molecularweight and composition. Lewis, “Controlled release of bioactive agentsfrom lactide/glycolide polymer,” in: M. Chasin and R. Langer (Eds.),“Biodegradable Polymers as Drug Delivery Systems (Marcel Dekker: NewYork, 1990), pp. 1-41.

[0737] This invention encompasses methods of screening compounds toidentify those that mimic the PRO polypeptide (agonists) or prevent theeffect of the PRO polypeptide (antagonists). Screening assays forantagonist drug candidates are designed to identify compounds that bindor complex with the PRO polypeptides encoded by the genes identifiedherein, or otherwise interfere with the interaction of the encodedpolypeptides with other cellular proteins. Such screening assays willinclude assays amenable to high-throughput screening of chemicallibraries, making them particularly suitable for identifying smallmolecule drug candidates.

[0738] The assays can be performed in a variety of formats, includingprotein-protein binding assays, biochemical screening assays,immunoassays, and cell-based assays, which are well characterized in theart.

[0739] All assays for antagonists are common in that they call forcontacting the drug candidate with a PRO polypeptide encoded by anucleic acid identified herein under conditions and for a timesufficient to allow these two components to interact.

[0740] In binding assays, the interaction is binding and the complexformed can be isolated or detected in the reaction mixture. In aparticular embodiment, the PRO polypeptide encoded by the geneidentified herein or the drug candidate is immobilized on a solid phase,e.g., on a microtiter plate, by covalent or non-covalent attachments.Non-covalent attachment generally is accomplished by coating the solidsurface with a solution of the PRO polypeptide and drying.Alternatively, an immobilized antibody, e.g., a monoclonal antibody,specific for the PRO polypeptidLe to be immobilized can be used toanchor it to a solid surface. The assay is performed by adding thenon-immobilized component, which may be labeled by a detectable label,to the immobilized component, e.g., the coated surface containing theanchored component. When the reaction is complete, the non-reactedcomponents are removed, e.g., by washing, and complexes anchored on thesolid surface are detected. When the originally non-immobilizedcomponent carries a detectable label, the detection of label immobilizedon the surface indicates that complexing occurred. Where the originallynon-immobilized component does not carry a label, complexing can bedetected, for example, by using a labeled antibody specifically bindingthe immobilized complex.

[0741] If the candidate compound interacts with but does not bind to aparticular PRO polypeptide encoded by a gene identified herein, itsinteraction with that polypeptide can be assayed by methods well knownfor detecting protein-protein interactions. Such assays includetraditional approaches, such as, e.g., cross-linking,co-immunoprecipitation, and co-purification through gradients orchromatographic columns. In addition, protein-protein interactions canbe monitored by using a yeast-based genetic system described by Fieldsand co-workers (Fields and Song, Nature (London), 340:245-246 (1989);Chien et al., Proc. Natl. Acad. Sci. USA, 88:9578-9582 (1991)) asdisclosed by Chevray and Nathans, Proc. Natl. Acad. Sci. USA, 89:5789-5793 (1991). Many transcriptional activators, such as yeast GAL4,consist of two physically discrete modular domains, one acting as theDNA-binding domain, the other one functioning as thetranscription-activation domain. The yeast expression system describedin the foregoing publications (generally referred to as the “two-hybridsystem”) takes advantage of this property, and employs two hybridproteins, one in which the target protein is fused to the DNA-bindingdomain of GAL4, and another, in which candidate activating proteins arefused to the activation domain. The expression of a GAL1-lacZ reportergene under control of a GAL4-activated promoter depends onreconstitution of GAL4 activity via protein-protein interaction.Colonies containing interacting polypeptides are detected with achromogenic substrate for β-galactosidase. A complete kit (MATCHMAKER™)for identifying protein-protein interactions between two specificproteins using the two-hybrid technique is commercially available fromClontech. This system can also be extended to map protein domainsinvolved in specific protein interactions as well as to pinpoint aminoacid residues that are crucial for these interactions.

[0742] Compounds that interfere with the interaction of a gene encodinga PRO polypeptide identified herein and other intra- or extracellularcomponents can be tested as follows: usually a reaction mixture isprepared containing the product of the gene and the intra- orextracellular component under conditions and for a time allowing for theinteraction and binding of the two products. To test the ability of acandidate compound to inhibit binding, the reaction is run in theabsence and in the presence of the test compound. In addition, a placebomay be added to a third reaction mixture, to serve as positive control.The binding (complex formation) between the test compound and the intra-or extracellular component present in the mixture is monitored asdescribed hereinabove. The formation of a complex in the controlreaction(s) but not in the reaction mixture containing the test compoundindicates that the test compound interferes with the interaction of thetest compound and its reaction partner.

[0743] To assay for antagonists, the PRO polypeptide may be added to acell along with the compound to be screened for a particular activityand the ability of the compound to inhibit the activity of interest inthe presence of the PRO polypeptide indicates that the compound is anantagonist to the PRO polypeptide. Alternatively, antagonists may bedetected by combining the PRO polypeptide and a potential antagonistwith membrane-bound PRO polypeptide receptors or recombinant receptorsunder appropriate conditions for a competitive inhibition assay. The PROpolypeptide can be labeled, such as by radioactivity, such that thenumber of PRO polypeptide molecules bound to the receptor can be used todetermine the effectiveness of the potential antagonist. The geneencoding the receptor can be identified by numerous methods known tothose of skill in the art, for example, ligand panning and FACS sorting.Coligan et al., Current Protocols in Immun., 1(2): Chapter 5 (1991).Preferably, expression cloning is employed wherein polyadenylated RNA isprepared from a cell responsive to the PRO polypeptide and a cDNAlibrary created from this RNA is divided into pools and used totransfect COS cells or other cells that are not responsive to the PROpolypeptide. Transfected cells that are grown on glass slides areexposed to labeled PRO polypeptide. The PRO polypeptide can be labeledby a variety of means including iodination or inclusion of a recognitionsite for a site-specific protein kinase. Following fixation andincubation, the slides are subjected to autoradiographic analysis.Positive pools are identified and sub-pools are prepared andre-transfected using an interactive sub-pooling and re-screeningprocess, eventually yielding a single clone that encodes the putativereceptor.

[0744] As an alternative approach for receptor identification, labeledPRO polypeptide can be photoaffinity-linked with cell membrane orextract preparations that express the receptor molecule. Cross-linkedmaterial is resolved by PAGE and exposed to X-ray film. The labeledcomplex containing the receptor can be excised, resolved into peptidefragments, and subjected to protein micro-sequencing. The amino acidsequence obtained from micro-sequencing would be used to design a set ofdegenerate oligonucleotide probes to screen a cDNA library to identifythe gene encoding the putative receptor. In another assay forantagonists, mammalian cells or a membrane preparation expressing thereceptor would be incubated with labeled PRO polypeptide in the presenceof the candidate compound. The ability of the compound to enhance orblock this interaction could then be measured.

[0745] More specific examples of potential antagonists include anoligonucleotide that binds to the fusions of immunoglobulin with PROpolypeptide, and, in particular, antibodies including, withoutlimitation, poly- and monoclonal antibodies and antibody fragments,single-chain antibodies, anti-idiotypic antibodies, and chimeric orhumanized versions of such antibodies or fragments, as well as humanantibodies and antibody fragments. Alternatively, a potential antagonistmay be a closely related protein, for example, a mutated form of the PROpolypeptide that recognizes the receptor but imparts no effect, therebycompetitively inhibiting the action of the PRO polypeptide.

[0746] Another potential PRO polypeptide antagonist is an antisense RNAor DNA construct prepared using antisense technology, where, e.g., anantisense RNA or DNA molecule acts to block directly the translation ofmRNA by hybridizing to targeted mRNA and preventing protein translation.Antisense technology can be used to control gene expression throughtriple-helix formation or antisense DNA or RNA, both of which methodsare based on binding of a polynucleotide to DNA or RNA. For example, the5′ coding portion of the polynucleotide sequence, which encodes themature PRO polypeptides herein, is used to design an antisense RNAoligonucleotide of from about 10 to 40 base pairs in length. A DNAoligonucleotide is designed to be complementary to a region of the geneinvolved in transcription (triple helix—see Lee et al., Nucl. AcidsRes., 6:3073 (1979); Cooney et al., Science, 241: 456 (1988); Dervan etal., Science, 251:1360 (1991)), thereby preventing transcription and theproduction of the PRO polypeptide. The antisense RNA oligonucleotidehybridizes to the mRNA in vivo and blocks translation of the mRNAmolecule into the PRO polypeptide (antisense—Okano, Neurochem., 56:560(1991); Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression(CRC Press: Boca Raton, Fla., 1988). The oligonucleotides describedabove can also be delivered to cells such that the antisense RNA or DNAmay be expressed in vivo to inhibit production of the PRO polypeptide.When antisense DNA is used, oligodeoxyribonucleotides derived from thetranslation-initiation site, e.g., between about −10 and +10 positionsof the target gene nucleotide sequence, are preferred.

[0747] Potential antagonists include small molecules that bind to theactive site, the receptor binding site, or growth factor or otherrelevant binding site of the PRO polypeptide, thereby blocking thenormal biological activity of the PRO polypeptide. Examples of smallmolecules include, but are not limited to, small peptides orpeptide-like molecules, preferably soluble peptides, and syntheticnon-peptidyl organic or inorganic compounds.

[0748] Ribozymes are enzymatic RNA molecules capable of catalyzing thespecific cleavage of RNA. Ribozymes act by sequence-specifichybridization to the complementary target RNA, followed byendonucleolytic cleavage. Specific ribozyme cleavage sites within apotential RNA target can be identified by known techniques. For furtherdetails see, e.g., Rossi, Current Biology, 4:469-471 (1994), and PCTpublication No. WO 97/33551 (published Sep. 18, 1997).

[0749] Nucleic acid molecules in triple-helix formation used to inhibittranscription should be single-stranded and composed ofdeoxynucleotides. The base composition of these oligonucleotides isdesigned such that it promotes triple-helix formation via Hoogsteenbase-pairing rules, which generally require sizeable stretches ofpurines or pyrimidines, on one strand of a duplex. For further detailssee, e.g., PCT publication No. WO 97/33551, supra.

[0750] These small molecules can be identified by any one or more of thescreening assays discussed hereinabove and/or by any other screeningtechniques well known for those skilled in the art.

[0751] Uses of the herein disclosed molecules may also be based upon thepositive functional assay hits disclosed and described below.

[0752] F. Anti-PRO Antibodies

[0753] The present invention further provides anti-PRO antibodies.Exemplary antibodies include polyclonal, monoclonal, humanized,bispecific, and heteroconjugate antibodies.

[0754] 1. Polyclonal Antibodies

[0755] The anti-PRO antibodies may comprise polyclonal antibodies.Methods of preparing polyclonal antibodies are known to the skilledartisan. Polyclonal antibodies can be raised in a mammal, for example,by one or more injections of an immunizing agent and, if desired, anadjuvant. Typically, the immunizing agent and/or adjuvant will beinjected in the mammal by multiple subcutaneous or intraperitonealinjections. The immunizing agent may include the PRO polypeptide or afusion protein thereof. It may be useful to conjugate the immunizingagent to a protein known to be immunogenic in the mammal beingimmunized. Examples of such immunogenic proteins include but are notlimited to keyhole limpet hemocyanin, serum albumin, bovinethyroglobulin, and soybean trypsin inhibitor. Examples of adjuvantswhich may be employed include Freund's complete adjuvant and MPL-TDMadjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).The immunization protocol may be selected by one skilled in the artwithout undue experimentation.

[0756] 2. Monoclonal Antibodies

[0757] The anti-PRO antibodies may, alternatively, be monoclonalantibodies. Monoclonal antibodies may be prepared using hybridomamethods, such as those described by Kohler and Milstein, Nature, 256:495(1975). In a hybridoma method, a mouse, hamster, or other appropriatehost animal, is typically immunized with an immunizing agent to elicitlymphocytes that produce or are capable of producing antibodies thatwill specifically bind to the immunizing agent. Alternatively, thelymphocytes may be immunized in vitro.

[0758] The immunizing agent will typically include the PRO polypeptideor a fusion protein thereof. Generally, either peripheral bloodlymphocytes (“PBLs”) are used if cells of human origin are desired, orspleen cells or lymph node cells are used if non-human mammalian sourcesare desired. The lymphocytes are then fused with an immortalized cellline using a suitable fusing agent, such as polyethylene glycol, to forma hybridoma cell [Goding, Monoclonal Antibodies: Principles andPractice, Academic Press, (1986) pp. 59-103]. Immortalized cell linesare usually transformed mammalian cells, particularly myeloma cells ofrodent, bovine and human origin. Usually, rat or mouse myeloma celllines are employed. The hybridoma cells may be cultured in a suitableculture medium that preferably contains one or more substances thatinhibit the growth or survival of the unfused, immortalized cells. Forexample, if the parental cells lack the enzyme hypoxanthine guaninephosphoribosyl transferase (HGPRT or HPRT), the culture medium for thehybridomas typically will include hypoxanthine, aminopterin, andthymidine (“HAT medium”), which substances prevent the growth ofHGPRT-deficient cells.

[0759] Preferred immortalized cell lines are those that fuseefficiently, support stable high level expression of antibody by theselected antibody-producing cells, and are sensitive to a medium such asHAT medium. More preferred immortalized cell lines are murine myelomalines, which can be obtained, for instance, from the Salk Institute CellDistribution Center, San Diego, Calif. and the American Type CultureCollection, Manassas, Va. Human myeloma and mouse-human heteromyelomacell lines also have been described for the production of humanmonoclonal antibodies [Kozbor, J. Immunol., 133:3001 (1984); Brodeur etal., Monoclonal Antibody Production Techniques and Applications, MarcelDekker, Inc., New York, (1987) pp. 51-63].

[0760] The culture medium in which the hybridoma cells are cultured canthen be assayed for the presence of monoclonal antibodies directedagainst PRO. Preferably, the binding specificity of monoclonalantibodies produced by the hybridoma cells is determined byimmunoprecipitation or by an in vitro binding assay, such asradioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchardanalysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0761] After the desired hybridoma cells are identified, the clones maybe subeloned by limiting dilution procedures and grown by standardmethods [Goding, supral. Suitable culture media for this purposeinclude, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640medium. Alternatively, the hybridoma cells may be grown in vivo asascites in a mammal.

[0762] The monoclonal antibodies secreted by the subclones may beisolated or purified from the culture medium or ascites fluid byconventional immunoglobulin purification procedures such as, forexample, protein A-Sepharose, hydroxylapatite chromatography, gelelectrophoresis, dialysis, or affinity chromatography.

[0763] The monoclonal antibodies may also be made by recombinant DNAmethods, such as those described in U.S. Pat. No. 4,816,567. DNAencoding the monoclonal antibodies of the invention can be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of murine antibodies). The hybridomacells of the invention serve as a preferred source of such DNA. Onceisolated, the DNA may be placed into expression vectors, which are thentransfected into host cells such as simian COS cells, Chinese hamsterovary (CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells. The DNA also may be modified, forexample, by substituting the coding sequence for human heavy and lightchain constant domains in place of the homologous murine sequences [U.S.Pat. No. 4,816,567; Morrison et al., supra] or by covalently joining tothe immunoglobulin coding sequence all or part of the coding sequencefor a non-immunoglobulin polypeptide. Such a non-immunoglobulinpolypeptide can be substituted for the constant domains of an antibodyof the invention, or can be substituted for the variable domains of oneantigen-combining site of an antibody of the invention to create achimeric bivalent antibody.

[0764] The antibodies may be monovalent antibodies. Methods forpreparing monovalent antibodies are well known in the art. For example,one method involves recombinant expression of immunoglobulin light chainand modified heavy chain. The heavy chain is truncated generally at anypoint in the Fc region so as to prevent heavy chain crosslinking.Alternatively, the relevant cysteine residues are substituted withanother amino acid residue or are deleted so as to prevent crosslinking.

[0765] In vitro methods are also suitable for preparing monovalentantibodies. Digestion of antibodies to produce fragments thereof,particularly, Fab fragments, can be accomplished using routinetechniques known in the art.

[0766] 3. Human and Humanized Antibodies

[0767] The anti-PRO antibodies of the invention may further comprisehumanized antibodies or human antibodies. Humanized forms of non-human(e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulinchains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)₂ or otherantigen-binding subsequences of antibodies) which contain minimalsequence derived from non-human immunoglobulin. Humanized antibodiesinclude human immunoglobulins (recipient antibody) in which residuesfrom a complementary determining region (CDR) of the recipient arereplaced by residues from a CDR of a non-human species (donor antibody)such as mouse, rat or rabbit having the desired specificity, affinityand capacity. In some instances, Fv framework residues of the humanimmunoglobulin are replaced by corresponding non-human residues.Humanized antibodies may also comprise residues which are found neitherin the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the CDR regions correspond to thoseof a non-human immunoglobulin and all or substantially all of the FRregions are those of a human immunoglobulin consensus sequence. Thehumanized antibody optimally also will comprise at least a portion of animmunoglobulin constant region (Fc), typically that of a humanimmunoglobulin [Jones et al., Nature, 321:522-525 (1986); Riechmann etal., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.,2:593-596 (1992)].

[0768] Methods for humanizing non-human antibodies are well known in theart. Generally, a humanized antibody has one or more amino acid residuesintroduced into it from a source which is non-human. These non-humanamino acid residues are often referred to as “import” residues, whichare typically taken from an “import” variable domain. Humanization canbe essentially performed following the method of Winter and co-workers[Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature,332:323-327 (1988); Verhoeyen et al., Science, 239:1534-15:36 (1988)],by substituting rodent CDRs or CDR sequences for the correspondingsequences of a human antibody. Accordingly, such “humanized” antibodiesare chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantiallyless than an intact human variable domain has been substituted by thecorresponding sequence from a non-human species. In practice, humanizedantibodies are typically human antibodies in which some CDR residues andpossibly some FR residues are substituted by residues from analogoussites in rodent antibodies.

[0769] Human antibodies can also be produced using various techniquesknown in the art, including phage display libraries [Hoogenboom andWinter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol.,222:581 (1991)]. The techniques of Cole et al. and Boemer et al. arealso available for the preparation of human monoclonal antibodies (Coleet al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77(1985) and Boemer et al., J. Immunol., 147(1):86-95 (1991)]. Similarly,human antibodies can be made by introducing of human immunoglobulin lociinto transgenic animals, e.g., nice in which the endogenousimmunoglobulin genes have been partially or completely inactivated. Uponchallenge, human antibody production is observed, which closelyresembles that seen in humans in all respects, including generearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the followingscientific publications: Marks et al., Bio/Technology 10, 779-783(1992); Lonberg etal., Nature 368 856-859 (1994); Morrison, Nature 368,812-13 (1994); Fishwild et al., Nature Biotechnology 14, 845-51 (1996);Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and [luszar,Intern. Rev. Immunol. 13 65-93 (1995).

[0770] 4. Bispecific Antibodies

[0771] Bispecific antibodies are monoclonal, preferably human orhumanized, antibodies that have binding specificities for at least twodifferent antigens. In the present case, one of the bindingspecificities is for the PRO, the other one is for any other antigen,and preferably for a cell-surface protein or receptor or receptorsubunit.

[0772] Methods for making bispecific antibodies are known in the art.Traditionally, the recombinant production of bispecific antibodies isbased on the co-expression of two immunoglobulin heavy-chain/light-chainpairs, where the two heavy chains have different specificities [Milsteinand Cuello, Nature, 305:537-539 (1983)]. Because of the randomassortment of immunoglobulin heavy and light chains, these hybridomas(quadromas) produce a potential mixture of ten different antibodymolecules, of which only one has the correct bispecific structure. Thepurification of the correct molecule is usually accomplished by affinitychromatography steps. Similar procedures are disclosed in WO 93/08829,published May 13, 1993, and in Traunecker et al., EMBO J, 10:3655-3659(1991).

[0773] Antibody variable domains with the desired binding specificities(antibody-antigen combining sites) can be fused to immunoglobulinconstant domain sequences. The fusion preferably is with animmunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the firstheavy-chain constant region (CH1) containing the site necessary forlight-chain binding present in at least one of the fusions. DNAsencoding the immunoglobulin heavy-chain fusions and, if desired, theimmunoglobulin light chain, are inserted into separate expressionvectors, and are co-transfected into a suitable host organism. Forfurther details of generating bispecific antibodies see, for example,Suresh et al., Methods in Enzymology, 121:210 (1986).

[0774] According to another approach described in WO 96/27011, theinterface between a pair of antibody molecules can be engineered tomaximize the percentage of heterodimers which are recovered fromrecombinant cell culture. The preferred interface comprises at least apart of the CH3 region of an antibody constant domain. In this method,one or more small amino acid side chains from the interface of the firstantibody molecule are replaced with larger side chains (e.g. tyrosine ortryptophan). Compensatory “cavities” of identical or similar size to thelarge side chain(s) are created on the interface of the second antibodymolecule by replacing large amino acid side chains with smaller ones(e.g. alanine or threonine). This provides a mechanism for increasingthe yield of the heterodimer over other unwanted end-products such ashomodimers.

[0775] Bispecific antibodies can be prepared as full length antibodiesor antibody fragments (e.g. F(ab′)₂ bispecific antibodies). Techniquesfor generating bispecific antibodies from antibody fragments have beendescribed in the literature. For example, bispecific antibodies can beprepared can be prepared using chemical linkage. Brennan et al., Science229:81 (1985) describe a procedure wherein intact antibodies areproteolytically cleaved to generate F(ab′)₂ fragments. These fragmentsare reduced in the presence of the dithiol complexing agent sodiumarsenite to stabilize vicinal dithiols and prevent intermoleculardisulfide formation. The Fab′ fragments generated are then converted tothionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives isthen reconverted to the Fab′-thiol by reduction with mercaptoethylamineand is mixed with an equimolar amount of the other Fab′-TNB derivativeto form the bispecific antibody. The bispecific antibodies produced canbe used as agents for the selective immobilization of enzymes.

[0776] Fab′ fragments may be directly recovered from E. coli andchemically coupled to form bispecific antibodies. Shalaby et al., J.Exp. Med. 175:217-225 (1992) describe the production of a fullyhumanized bispecific antibody F(ab′)₂ molecule. Each Fab′ fragment wasseparately secreted from E. coli and subjected to directed chemicalcoupling in vitro to form the bispecific antibody. The bispecificantibody thus formed was able to bind to cells overexpressing the ErbB2receptor and normal human T cells, as well as trigger the lytic activityof human cytotoxic lymphocytes against human breast tumor targets.

[0777] Various technique for making and isolating bispecific antibodyfragments directly from recombinant cell culture have also beendescribed. For example, bispecific antibodies have been produced usingleucine zippers. Kostelny et al., J. Immunol. 148(5): 1547-1553 (1992).The leucine zipper peptides from the Fos and Jun proteins were linked tothe Fab′ portions of two different antibodies by gene fusion. Theantibody homodimers were reduced at the hinge region to form monomersand then re-oxidized to form the antibody heterodimers. This method canalso be utilized for the production of antibody homodimers. The“diabody” technology described by Hollinger et al., Proc. Natl. Acad.Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism formaking bispecific antibody fragments. The fragments comprise aheavy-chain variable domain (V_(H)) connected to a light-chain variabledomain (V_(L)) by a linker which is too short to allow pairing betweenthe two domains on the same chain. Accordingly, the V_(H) and V_(L)domains of one fragment are forced to pair with the complementary V_(L)and V_(H) domains of another fragment, thereby forming twoantigen-binding sites. Another strategy for making bispecific antibodyfragments by the use of single-chain Fv (sFv) dimers has also beenreported. See, Gruber et al., J. Immunol. 152:5368 (1994). Antibodieswith more than two valencies are contemplated. For example, trispecificantibodies can be prepared. Tutt el al., J. Immunol. 147:60 (1991).

[0778] Exemplary bispecific antibodies may bind to two differentepitopes on a given PRO polypeptide herein. Alternatively, an anti-PROpolypeptide arm may be combined with an arm which binds to a triggeringmolecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2,CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64),FcγRII (CD32) and FcγRIII (CD16) so as to focus cellular defensemechanisms to the cell expressing the particular PRO polypeptide.Bispecific antibodies may also be used to localize cytotoxic agents tocells which express a particular PRO polypeptide. These antibodiespossess a PRO-binding arm and an arm which binds a cytotoxic agent or aradionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Anotherbispecific antibody of interest binds the PRO polypeptide and furtherbinds tissue factor (TF).

[0779] 5.Heteroconiugate Antibodies

[0780] Heteroconjugate antibodies are also within the scope of thepresent invention. Heteroconjugate antibodies are composed of twocovalentlyjoined antibodies. Such antibodies have, for example, beenproposed to target immune system cells to unwanted cells [U.S. Pat. No.4,676,980], and for treatment of HIV infection [WO 91/00360; WO92/200373; EP 03089]. It is contemplated that the antibodies may beprepared in vitro using known methods in synthetic protein chemistry,including those involving crosslinking agents. For example, immunotoxinsmay be constructed using a disulfide exchange reaction or by forming athioether bond. Examples of suitable reagents for this purpose includeiminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, forexample, in U.S. Pat. No. 4,676,980.

[0781] 6. Effector Function Engineering

[0782] It may be desirable to modify the antibody of the invention withrespect to effector function, so as to enhance, e.g., the effectivenessof the antibody in treating cancer. For example, cysteine residue(s) maybe introduced into the Fc region, thereby allowing interchain disulfidebond formation in this region. The homodimeric antibody thus generatedmay have improved internalization capability and/or increasedcomplement-mediated cell killing and antibody-dependent cellularcytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195(1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimericantibodies with enhanced anti-tumor activity may also be prepared usingheterobifunctional cross-linkers as described in Wolff et al. CancerResearch, 53: 2560-2565 (1993). Alternatively, an antibody can beengineered that has dual Fc regions and may thereby have enhancedcomplement lysis and ADCC capabilities. See Stevenson et al.,Anti-Cancer Drug Design. 3: 219-230 (1989).

[0783] 7. Immunoconjugates

[0784] The invention also pertains to immunoconjugates comprising anantibody conjugated to a cytotoxic agent such as a chemotherapeuticagent, toxin (e.g., an enzymatically active toxin of bacterial, fungal,plant, or animal origin, or fragments thereof), or a radioactive isotope(i.e., a radioconjugate).

[0785] Chemotherapeutic agents useful in the generation of suchimmunoconjugates have been described above. Enzymatically active toxinsand fragments thereof that can be used include diphtheria A chain,nonbinding active fragments of diphtheria toxin, exotoxin A chain (fromPseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain,alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolacaamericana proteins (PAPI, PAPII, and PAP-S), momordica charantiainhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin,mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. Avariety of radionuclides are available for the production ofradioconjugated antibodies. Examples include ²¹²Bi, t ¹³¹In, ⁹⁰Y, and¹⁸⁶Re.

[0786] Conjugates of the antibody and cytotoxic agent are made using avariety of bifunctional protein-coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glulareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin canbe prepared as described in Vitetta et al., Science, 238: 1098 (1987).Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. See WO94/11026.

[0787] In another embodiment, the antibody may be conjugated to a“receptor” (such streptavidin) for utilization in tumor pretargetingwherein the antibody-receptor conjugate is administered to the patient,followed by removal of unbound conjugate from the circulation using aclearing agent and then administration of a “ligand” (e.g., avidin) thatis conjugated to a cytotoxic agent (e.g., a radionucleotide).

[0788] 8. Immunoliposomes

[0789] The antibodies disclosed herein may also be formulated asimmunoliposomes. Liposomes containing the antibody are prepared bymethods known in the art, such as described in Epstein et al., Proc.Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl Acad.Sci. USA, 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545.Liposomes with enhanced circulation time are disclosed in U.S. Pat. No.5,013,556.

[0790] Particularly useful liposomes can be generated by thereverse-phase evaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol, and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter. Fab′ fragments of the antibody of the present invention can beconjugated to the liposomes as described in Martin et al., J. Biol.Chem., 257: 286-288 (1982) via a disulfide-interchange reaction. Achemotherapeutic agent (such as Doxorubicin) is optionally containedwithin the liposome. See Gabizon et al., J. National Cancer Inst.,81(19): 1484 (1989).

[0791] 9. Pharmaceutical Compositions of Antibodies

[0792] Antibodies specifically binding a PRO polypeptide identifiedherein, as well as other molecules identified by the screening assaysdisclosed hereinbefore, can be administered for the treatment of variousdisorders in the form of pharmaceutical compositions.

[0793] If the PRO polypeptide is intracellular and whole antibodies areused as inhibitors, internalizing antibodies are preferred. However,lipofections or liposomes can also be used to deliver the antibody, oran antibody fragment, into cells. Where antibody fragments are used, thesmallest inhibitory fragment that specifically binds to the bindingdomain of the target protein is preferred. For example, based upon thevariable-region sequences of an antibody, peptide molecules can bedesigned that retain the ability to bind the target protein sequence.Such peptides can be synthesized chemically and/or produced byrecombinant DNA technology. See, e.g., Marasco et al., Proc. Natl. Acad.Sci. USA, 90: 7889-7893 (1993). The formation herein may also containmore than one active compound as necessary for the particular indicationbeing treated, preferably those with complementary activities that donot adversely affect each other. Alternatively, or in addition, thecomposition may comprise an agent that enhances its function, such as,for example, a cytotoxic agent, cytokine, chemotherapeutic agent, orgrowth-inhibitory agent. Such molecules are suitably present incombination in amounts that are effective for the purpose intended.

[0794] The active ingredients may also be entrapped in microcapsulesprepared, for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles, andnanocapsules) or in macroemulsions. Such techniques are disclosed inRemington's Pharmaceutical Sciences, supra.

[0795] The formulations to be used for in vivo administration must besterile. This is readily accomplished by filtration through sterilefiltration membranes.

[0796] Sustained-release preparations may be prepared. Suitable examplesof sustained-release preparations include semipermeable matrices ofsolid hydrophobic polymers containing the antibody, which matrices arein the form of shaped articles, e.g., films, or microcapsules. Examplesof sustained-release matrices include polyesters, hydrogels (forexample, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate,degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid.While polymers such as ethylene-vinyl acetate and lactic acid-glycolicacid enable release of molecules for over 100 days, certain hydrogelsrelease proteins for shorter time periods. When encapsulated antibodiesremain in the body for a long time, they may denature or aggregate as aresult of exposure to moisture at 37° C., resulting in a loss ofbiological activity and possible changes in immunogenicity. Rationalstrategies can be devised for stabilization depending on the mechanisminvolved. For example, if the aggregation mechanism is discovered to beintermolecular S—S bond formation through thio-disulfide interchange,stabilization may be achieved by modifying sulfhydryl residues,lyophilizing from acidic solutions, controlling moisture content, usingappropriate additives, and developing specific polymer matrixcompositions.

[0797] G. Uses for anti-PRO Antibodies

[0798] The anti-PRO antibodies of the invention have various utilities.For example, anti-PRO antibodies may be used in diagnostic assays forPRO, e.g., detecting its expression in specific cells, tissues, orserum. Various diagnostic assay techniques known in the art may be used,such as competitive binding assays, direct or indirect sandwich assaysand immunoprecipitation assays conducted in either heterogeneous orhomogeneous phases [Zola, Monoclonal Antibodies: A Manual of Techniques,CRC Press, Inc. (1987) pp. 147-158]. The antibodies used in thediagnostic assays can be labeled with a detectable moiety. Thedetectable moiety should be capable of producing, either directly orindirectly, a detectable signal. For example, the detectable moiety maybe a radioisotope, such as ³H, ¹⁴C, ³²P, ³⁵S, or ¹²⁵I, a fluorescent orchemiluminescent compound, such as fluorescein isothiocyanate,rhodamine, or luciferin, or an enzyme, such as alkaline phosphatase,beta-galactosidase or horseradish peroxidase. Any method known in theart for conjugating the antibody to the detectable moiety may beemployed, including those methods described by Hunter et al., Nature,144:945 (1962); David et al., Biochemistry, 13:1014 (1974); Pain et al.,J. Immunol. Meth., 40:219 (1981); and Nygren, J. Histochem. andCytochem., 30:407 (1982).

[0799] Anti-PRO antibodies also are useful for the affinity purificationof PRO from recombinant cell culture or natural sources. In thisprocess, the antibodies against PRO are immobilized on a suitablesupport, such a Sephadex resin or filter paper, using methods well knownin the art. The immobilized antibody then is contacted with a samplecontaining the PRO to be purified, and thereafter the support is washedwith a suitable solvent that will remove substantially all the materialin the sample except the PRO, which is bound to the immobilizedantibody. Finally, the support is washed with another suitable solventthat will release the PRO from the antibody.

[0800] The following examples are offered for illustrative purposesonly, and are not intended to limit the scope of the present inventionin any way.

[0801] All patent and literature references cited in the presentspecification are hereby incorporated by reference in their entirety.

EXAMPLES

[0802] Commercially available reagents referred to in the examples wereused according to manufacturer's instructions unless otherwiseindicated. The source of those cells identified in the followingexamples, and throughout the specification, by ATCC accession numbers isthe American Type Culture Collection, Manassas, Va.

Example 1 Extracellular Domain Homology Screening to Identify NovelPolypeptides and cDNA Encoding Therefor

[0803] The extracellular domain (ECD) sequences (including the secretionsignal sequence, if any) from about 950 known secreted proteins from theSwiss-Prot public database were used to search EST databases. The ESTdatabases included public databases (e.g., Dayhoff, GenBank), andproprietary databases (e.g. LIFESEQ™, Incyte Pharmaceuticals, Palo Alto,Calif.). The search was performed using the computer program BLAST orBLAST-2 (Altschul et al., Methods in Enzymology 266:460-480 (1996)) as acomparison of the ECD protein sequences to a 6 frame translation of theEST sequences. Those comparisons with a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into consensus DNA sequences with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.).

[0804] Using this extracellular domain homology screen, consensus DNAsequences were assembled relative to the other identified EST sequencesusing phrap. In addition, the consensus DNA sequences obtained wereoften (but not always) extended using repeated cycles of BLAST orBLAST-2 and phrap to extend the consensus sequence as far as possibleusing the sources of EST sequences discussed above.

[0805] Based upon the consensus sequences obtained as described above,oligonucleotides were then synthesized and used to identify by PCR acDNA library that contained the sequence of interest and for use asprobes to isolate a clone of the full-length coding sequence for a PROpolypeptide. Forward and reverse PCR primers generally range from 20 to30 nucleotides and are often designed to give a PCR product of about100-1000 bp in length. The probe sequences are typically 40-55 bp inlength. In some cases, additional oligonucleotides are synthesized whenthe consensus sequence is greater than about 1-1.5kbp. In order toscreen several libraries for a full-length clone, DNA from the librarieswas screened by PCR amplification, as per Ausubel et al., CurrentProtocols in Molecular Biology, with the PCR primer pair. A positivelibrary was then used to isolate clones encoding the gene of interestusing the probe oligonucleotide and one of the primer pairs.

[0806] The cDNA libraries used to isolate the cDNA clones wereconstructed by standard methods using commercially available reagentssuch as those from Invitrogen, San Diego, Calif. The cDNA was primedwith oligo dT containing a NotI site, linked with blunt to SalIhemikinased adaptors, cleaved with NotI, sized appropriately by gelelectrophoresis, and cloned in a defined orientation into a suitablecloning vector (such as pRKB or pRKD; pRK5B is a precursor of pRK5D thatdoes not contain the SfiI site; see, Holmes et al., Science,253:1278-1280 (1991),) inthe unique XhoI and NotI sites.

Example 2 Isolation of cDNA clones by Amylase Screening

[0807] 1. Preparation of oligo dT primed cDNA library

[0808] mRNA was isolated from a human tissue of interest using reagentsand protocols from Invitrogen, San Diego, Calif. (Fast Track 2). ThisRNA was used to generate an oligo dT primed cDNA library in the vectorpRK5D using reagents and protocols from Life Technologies, Gaithersburg,Md. (Super Script Plasmid System). In this procedure, the doublestranded cDNA was sized to greater than 1000 bp and the SalI/NotITinkered cDNA was cloned into XhoI/NotI cleaved vector. pRK5D is acloning vector that has an sp6 transcription initiation site followed byan Sfil restriction enzyme site preceding the XhoI/NotI cDNA cloningsites.

[0809] 2. Preparation of Random Primed cDNA Library

[0810] A secondary cDNA library was generated in order to preferentiallyrepresent the 5′ ends of the primary cDNA clones. Sp6 RNA was generatedfrom the primary library (described above), and this RNA was used togenerate a random primed cDNA library in the vector pSST-AMY.0 usingreagents and protocols from Life Technologies (Super Script PlasmidSystem, referenced above). In this procedure the double stranded cDNAwas sized to 500-1000 bp, Tinkered with blunt to NotI adaptors, cleavedwith SfiI, and cloned into SfiI/NotI cleaved vector. pSST-AMY.0 is acloning vector that has a yeast alcohol dehydrogenase promoter precedingthe cDNA cloning sites and the mouse amylase sequence (the maturesequence without the secretion signal) followed by the yeast alcoholdehydrogenase terminator, after the cloning sites. Thus, cDNAs clonedinto this vector that are fused in frame with amylase sequence will leadto the secretion of amylase from appropriately transfected yeastcolonies.

[0811] 3. Transformation and Detection

[0812] DNA from the library described in paragraph 2 above was chilledon ice to which was added electrocompetent DH10B bacteria (LifeTechnologies, 20 ml). The bacteria and vector mixture was thenelectroporated as recommended by the manufacturer. Subsequently, SOCmedia (Life Technologies, 1 ml) was added and the mixture was incubatedat 37° C. for 30 minutes. The transformants were then plated onto 20standard 150 mm LB plates containing ampicillin and incubated for 16hours (37° C.). Positive colonies were scraped off the plates and theDNA was isolated from the bacterial pellet using standard protocols,e.g. CsCl-gradient. The purified DNA was then carried on to the yeastprotocols below.

[0813] The yeast methods were divided into three categories: (1)Transformation of yeast with the plasmid/cDNA combined vector; (2)Detection and isolation of yeast clones secreting amylase; and (3) PCRamplification of the insert directly from the yeast colony andpurification of the DNA for sequencing and further analysis.

[0814] The yeast strain used was HD56-5A (ATCC-90785). This strain hasthe following genotype: MAT alpha, ura3-52, leu2-3, leu2-112, his3-11,his3-15, MAL⁺, SUC⁺, GAL⁺. Preferably, yeast mutants can be employedthat have deficient post-translational pathways. Such mutants may havetranslocation deficient alleles in sec71, sec72, sec62, with truncatedsec71 being most preferred. Alternatively, antagonists (includingantisense nucleotides and/or ligands) which interfere with the normaloperation of these genes, other proteins implicated in this posttranslation pathway (e.g., SEC61p, SEC72p, SEC62p, SEC63p, TDJlp orSSA1p-4p) or the complex formation of these proteins may also bepreferably employed in combination with the amylase-expressing yeast.

[0815] Transformation was performed based on the protocol outlined byGietz et al., Nucl. Acid. Res., 20:1425 (1992). Transformed cells werethen inoculated from agar into YEPD complex media broth (100 ml) andgrown overnight at 30° C. The YEPD broth was prepared as described inKaiser et al., Methods in Yeast Genetics, Cold Spring Harbor Press, ColdSpring Harbor, N.Y., p. 207 (1994). The overnight culture was thendiluted to about 2×10⁶ cells/ml (approx. OD₆₀₀=0.1) into fresh YEPDbroth (500 ml) and regrown to 1×10⁷ cells/ml (approx. OD₆=0.4−0.5).

[0816] The cells were then harvested and prepared for transformation bytransfer into GS3 rotor bottles in a Sorval GS3 rotor at 5,000 rpm for 5minutes, the supernatant discarded, and then resuspended into sterilewater, and centrifuged again in 50 ml falcon tubes at 3,500 rpm in aBeckman GS-6KR centrifuge. The supernatant was discarded and the cellswere subsequently washed with LiAc/TE (10 ml, 10 mM Tris-HCl, 1 mM EDTApH 7.5, 100 mM Li₂OOCCH₃), and resuspended into LiAc/TE (2.5 ml).

[0817] Transformation took place by mixing the prepared cells (100 μl)with freshly denatured single stranded salmon testes DNA (LofstrandLabs, Gaithersburg, Md.) and transforming DNA (1 μg, vol. <10 μl) inmicrofuge tubes. The mixture was mixed briefly by vortexing, then 40%PEG/TE (600 μl, 40% polyethylene glycol-4000, 10 mM Tris-HCl, 1 mM EDTA,100 mM Li₂OOCCH₃, pH 7.5) was added. This mixture was gently mixed andincubated at 30° C. while agitating for 30 minutes. The cells were thenheat shocked at 42° C. for 15 minutes, and the reactionvesselcentrifuged in a microfuge at 12,000 rpm for 5-10 seconds, decanted andresuspended into TE (500 μl, 10 mM Tris-HCl, 1 mM EDTA pH 7.5) followedby recentrifugation. The cells were then diluted into TE (1 ml) andaliquots (200 μl) were spread onto the selective media previouslyprepared in 150 mm growth plates (VWR).

[0818] Alternatively, instead of multiple small reactions, thetransformation was performed using a single, large scale reaction,wherein reagent amounts were scaled up accordingly.

[0819] The selective media used was a synthetic complete dextrose agarlacking uracil (SCD-Ura) prepared as described in Kaiser et al., Methodsin Yeast Genetics, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.,p. 208-210 (1994). Transformants were grown at 30° C. for 2-3 days.

[0820] The detection of colonies secreting amylase was performed byincluding red starch in the selective growth media. Starch was coupledto the red dye (Reactive Red-120, Sigma) as per the procedure describedby Biely et al., Anal. Biochem., 172:176-179 (1988). The coupled starchwas incorporated into the SCD-Ura agar plates at a final concentrationof 0.15% (w/v), and was buffered with potassium phosphate to a pH of 7.0(50-100 mM final concentration).

[0821] The positive colonies were picked and streaked across freshselective media (onto 150 mm plates) in order to obtain well isolatedand identifiable single colonies. Well isolated single colonies positivefor amylase secretion were detected by direct incorporation of redstarch into buffered SCD-Ura agar. Positive colonies were determined bytheir ability to break down starch resulting in a clear halo around thepositive colony visualized directly.

[0822] 4. Isolation of DNA by PCR Amplification

[0823] When a positive colony was isolated, a portion of it was pickedby a toothpick and diluted into sterile water (30 μl) in a 96 wellplate. At this time, the positive colonies were either frozen and storedfor subsequent analysis or immediately amplified. An aliquot of cells (5μl) was used as a template for the PCR reaction in a 25 μl volumecontaining: 0.5 μl Klentaq (Clontech, Palo Alto, Calif.); 4.0 μl 10 mMdNTP's (Perkin Elmer-Cetus); 2.5 μl Kentaq buffer (Clontech); 0.25 μlforward oligo 1; 0.25 reverse oligo 2; 12.5 μl distilled water.

[0824] The sequence of the forward oligonucleotide 1 was:

[0825] 5′-TGTAAAACGACGGCCAGTTAAATAGACCTGCAATTATTAATCT-3′ (SEQ ID NO:3)

[0826] The sequence of reverse oligonucleotide 2 was:

[0827] 5′-CAGGAAACAGCTATGACCACCTGCACACCTGCAAATCCATT-3′ (SEQ ID NO:4)

[0828] PCR was then performed as follows: a. Denature 92° C., 5 minutesb. 3 cycles of: Denature 92° C., 30 seconds Anneal 59° C., 30 secondsExtend 72° C., 60 seconds c. 3 cycles of: Denature 92° C., 30 secondsAnneal 57° C., 30 seconds Extend 72° C., 60 seconds d. 25 cycles of:Denature 92° C., 30 seconds Anneal 55° C., 30 seconds Extend 72° C., 60seconds e. Hold  4° C.

[0829] The underlined regions of the oligonucleotides annealed to theADH promoter region and the amylase region, respectively, and amplifieda 307 bp region from vector pSST-AMY.0 when no insert was present.Typically, the first 18 nucleotides of the 5′ end of theseoligonucleotides contained annealing sites for the sequencing primers.Thus, the total product of the PCR reaction from an empty vector was 343bp. However, signal sequence-fused-cDNA resulted in considerably longernucleotide sequences.

[0830] Following the PCR, an aliquot of the reaction (5 μl) was examinedby agarose gel electrophoresis in a 1% agarose gel using aTris-Borate-EDTA (TBE) buffering system as described by Sambrook et al.,supra. Clones resulting in a single strong PCR product larger than 400bp were further analyzed by DNA sequencing after purification with a 96Qiaquick PCR clean-up column (Qiagen Inc., Chatsworth, Calif.).

Example 3 Isolation of cDNA Clones Using Siznal Algorithm Analysis

[0831] Various polypeptide-encoding nucleic acid sequences wereidentified by applying a proprietary signal sequence finding algorithmdeveloped by Genentech, Inc. (South San Francisco, Calif.) upon ESTs aswell as clustered and assembled EST fragments from public (e.g.,GenBank) and/or private (LIFESEQ®, Incyte Pharmaceuticals, Inc., PaloAlto, Calif.) databases. The signal sequence algorithm computes asecretion signal score based on the character of the DNA nucleotidessurrounding the first and optionally the second methionine codon(s)(ATG) at the 5′-end of the sequence or sequence fragment underconsideration. The nucleotides following the first ATG must code for atleast 35 unambiguous amino acids without any stop codons. If the firstATG has the required amino acids, the second is not examined. If neithermeets the requirement, the candidate sequence is not scored. In order todetermine whether the EST sequence contains an authentic signalsequence, the DNA and corresponding amino acid sequences surrounding theATG codon are scored using a set of seven sensors (evaluationparameters) known to be associated with secretion signals. Use of thisalgorithm resulted in the identification of numerouspolypeptide-encoding nucleic acid sequences.

Example 4 Isolation of cDNA clones Encoding Human PRO1484

[0832] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA39616. Based on the DNA39616 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO1484.

[0833] PCR primers (forward and reverse) were synthesized: forward PCRprimer (39616.f1) 5′-GCAACAATGGAGCCACTGGTCATG-3′ (SEQ ID NO:5) reversePCR primer (39616.r1) 5′-GCAAAGGTGGAGAAGCGTTGGTGG-3′ (SEQ ID NO:6)

[0834] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39616 sequence which had the followingnucleotide sequence

[0835] hybridization probe (39616.p1)

[0836] 5′-CCCACTTCAGCAATCAGAACAGTGGGATTATCTTTCAGCAGTGTTTGAGACC-3′ (SEQID NO:7)

[0837] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1484 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidneytissue.

[0838] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1484 (designated herein as DNA44686-1653[FIG. 1, SEQ ID NO:1]; (UNQ753) and the derived protein sequence forPRO1484.

[0839] The entire nucleotide sequence of DNA44686-1653 is shown in FIG.1 (SEQ ID NO:1). Clone DNA44686-1653 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 77-79 and ending at the stop codon at nucleotide positions815-817 (FIG. 1). The predicted polypeptide precursor is 246 amino acidslong (FIG. 2). The full-length PRO1484 protein shown in FIG. 2 has anestimated molecular weight of about 26,994 daltons and a pI of about6.43. Analysis of the full-length PRO1484 sequence shown in FIG. 2 (SEQID NO:2) evidences the presence of the following: a signal peptide fromabout amino acid 1 to about amino acid 22, a Clq domain signaturesequence from about amino acid 137 to about amino acid 167 and variousamino acid sequence blocks having homology to Clq domain-containingproteins as shown in FIG. 2. Clone DNA44686-1653 has been deposited withATCC on Jan. 12, 1999 and is assigned ATCC deposit no. 203581.

[0840] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 2 (SEQ ID NO:2), evidenced significant homologybetween the PRO1484 amino acid sequence and the following Dayhoffsequences: P_WO9108, CA1A_HUMAN, C1QC_HUMAN, HUMC1QB2_(—)1, COLE_LEPMA,MMU32107_(—)1, CAS4_EPHMU, A57131, A41207 and CERL_RAT.

Example 5 Isolation of cDNA clones Encoding Human PRO4334

[0841] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA56421.

[0842] In light of an observed sequence homology between the DNA56421sequence and an EST sequence contained within the Incyte EST clone no.3347532, the Incyte clone was purchased and the cDNA insert was obtainedand sequenced. The sequence of this cDNA insert is shown in FIG. 3 andis herein designated as DNA59608-2577.

[0843] The full length clone shown in FIG. 3 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 83-85 and ending at the stop codon found atnucleotide positions 1404-1406 (FIG. 3; SEQ ID NO:8). The predictedpolypeptide precursor (FIG. 4, SEQ ID NO:9) is 440 amino acids long.PRO4334 has a calculated molecular weight of approximately 50,211daltons and an estimated pI of approximately 8.29.

[0844] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 4 (SEQ ID NO:9), revealed homology between thePRO4334 amino acid sequence and the following Dayhoff sequencesincorporated herein: AB020686_(—)1, PC1_HUMAN, P_R79148, PC1_MOUSE,RNU78788_(—)1, RATPDIB_(—)1, P_W75859, AC005587_(—)1, P_R86595 andPPD1_BOVIN.

[0845] Clone DNA5,9608-2577 was deposited with the ATCC on Mar. 23, 1999and is assigned ATCC deposit no. 203870.

Example 6 Isolation of cDNA clones Encoding Human PRO1122

[0846] An expressed sequence tag (EST) DNA database (LIFESEQ”, IncytePharmaceuticals, Palo Alto, Calif.) was searched and an EST wasidentified. The EST was Incyte 1347523 which is also called hereinDNA49665. Based on the DNA49665 sequence, oligonucleotides weresynthesized: 1) to identify by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence for PRO1122.

[0847] PCR primers (forward and reverse) were synthesized: forward PCRprimer 5′-ATCCACAGAAGCTGGCCTTCGCCG-3′; and (SEQ ID NO:12) reverse PCRprimer 5′-GGGACGTGGATGAACTCGGTGTGG-3′. (SEQ ID NO:13)

[0848] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed which had the following nucleotide sequence:

[0849] hybridization probe

[0850] 5′-TATCCACAGAAGCTGGCCTTCGCCGAGTGCCTGTGCAGAG-3′ (SEQ ID NO: 14)

[0851] In order to screen several libraries for a source of a fll-lengthclone, DNA from the libraries was screened by PCR amplification with thePCR primer pair identified above. A positive library was then used toisolate clones encoding the PRO1122 gene using the probe oligonucleotideand one of the PCR primers. RNA for construction of the cDNA librarieswas isolated from human fetal kidney tissue (LIB228).

[0852] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1122 [herein designated asDNA62377-1381] (SEQ ID NO:10) and the derived protein sequence forPRO1122.

[0853] The entire nucleotide sequence of DNA62377-1381 is shown in FIG.5 (SEQ ID NO:10). Clone DNA62377-1381 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 50-52 and ending at the stop codon at nucleotide positions641-643 of SEQ ID NO:10 (FIG. 5). The predicted polypeptide precursor is197 amino acids long (FIG. 6). The full-length PRO1122 protein shown inFIG. 6 has an estimated molecular weight of about 21,765 daltons and apI of about 8.53. Clone DNA62377-1381 has been deposited with ATCC onDec. 22, 1998. It is understood that the deposited clone has the actualnucleic acid sequence and that the sequences provided herein are basedon known sequencing techniques.

[0854] Analysis of the amino acid sequence of the full-length PRO1122polypeptide suggests that it possesses similarity with CTLA-8 and IL-17,thereby indicating that PRO1122 may be a novel cytokine. Morespecifically, an analysis of the Dayhoff database (version 35.45SwissProt 35) evidenced significant homology between the PRO1122 aminoacid sequence and the following Dayhoff sequences, P-W13651, VG13_HSVSAand CEF25D1_(—)1.

Example 7 Isolation of cDNA clones Encoding Human PRO1889

[0855] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA49310. Based up an observed homologybetween the DNA49310 consensus sequence and an EST contained within theIncyte EST clone no. 2779436, Incyte EST clone no. 2779436 was purchasedand its insert obtained and sequenced. The sequence of that insert isshown in FIG. 7 and is herein designated DNA77623-2524.

[0856] The entire nucleotide sequence of DNA77623-2524 is shown in FIG.7 (SEQ ID NO:15). Clone DNA77623-2524 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 39-41 and ending at the stop codon at nucleotide positions330-332 (FIG. 7). The predicted polypeptide precursor is 97 amino acidslong (FIG. 8). The full-length PRO1889 protein shown in FIG. 8 has anestimated molecular weight of about 10,160 daltons and a pI of about6.56. Analysis of the full-length PRO1889 sequence shown in FIG. 8 (SEQID NO:16) evidences the presence of the following: a signal peptide fromabout amino acid 1 to about amino acid 20, potential N-myristolationsites from about amino acid 6 to about amino acid 11 and from aboutamino acid 33 to about amino acid 38 and prokaryotic membranelipoprotein lipid attachment sites from about amino acid 24 to aboutamino acid 34 and from about amino acid 78 to about amino acid 88. CloneDNA77623-2524 has been deposited with ATCC on Dec. 22, 1998 and isassigned ATCC deposit no. 203546.

[0857] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 8 (SEQ ID NO:16), evidenced significant homologybetween the PRO1889 amino acid sequence and the following Dayhoffsequences: HSE48ATGN_(—)1, P_WO6292, AB012293_(—)1, THYB_MOUSE,P_R70984, CHKSCA2A_(—)1, P_W61628, I48639, BMBUNGKP4_(—)1 andUPAR_HUMAN.

Example 8 Isolation of cDNA clones Encoding Human PRO1890

[0858] A consensus DNA sequence was assembled relative to other ESTsequences using repeated cycles of BLAST and phrap as described inExample 1 above. This consensus sequence is herein designated DNA52162.Based on the DNA52162 consensus sequence, oligonucleotides weresynthesized: 1) to identify by PCR a cDNA library that contained thesequence of interest, and 2) for use as probes to isolate a clone of thefull-length coding sequence for PRO1890.

[0859] PCR primers (forward and reverse) were synthesized: forward PCRprimer (52162.f1) 5′-CACCAACCAACTGCCAATCCTGGC-3′ (SEQ ID NO:19) reversePCR primer (52162.r1) 5′-ACCACATTCTGATGGGTGTCTCCTGG-3′ (SEQ ID NO:20)

[0860] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA52162 sequence which had the followingnucleotide sequence

[0861] hybridization probe (52162.p1)

[0862] 5 ′-GGGTCCCTACCTTTACCAGTGGAATGATGACAGGTGTAACATGAAGCAC-3′ (SEQ IDNO:21)

[0863] RNA for construction of the cDNA libraries was isolated fromhuman fetal kidney tissue.

[0864] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1890 (designated herein as DNA79230-2525[FIG. 9, SEQ ID NO:17]; (UNQ872) and the derived protein sequence forPRO1890.

[0865] The entire nucleotide sequence of DNA79230-2525 is shown in FIG.9 (SEQ ID NO:17). Clone DNA79230-2525 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 378-380 and ending at the stop codon at nucleotide positions1197-1199 (FIG. 9). The predicted polypeptide precursor is 273 aminoacids long (FIG. 10). The full-length PRO1890 protein shown in FIG. 10has an estimated molecular weight of about 30,431 daltons and a pI ofabout 6.79. Analysis of the full-length PRO1890 sequence shown in FIG.10 (SEQ ID NO:18) evidences the presence of the following: a signalpeptide from about amino acid 1 to about amino acid 21, a transmembranedomain from about amino acid 214 to about amino acid 235, potentialN-glycosylation sites from about amino acid 86 to about amino acid 89and from about amino acid 255 to about amino acid 258, a cAMP- andcGMP-dependent protein kinase phosphorylation site from about amino acid266 to about amino acid 269 and potential N-myristolation sites fromabout amino acid 27 to about amino acid 32, from about amino acid 66 toabout amino acid 71, from about amino acid 91 to about amino acid 96,from about amino acid 93 to about amino acid 98, from about amino acid102 to about amino acid 107, from about amino acid 109 to about aminoacid 114, from about amino acid 140 to about amino acid 145 and fromabout amino acid 212 to about amino acid 217. Clone DNA79230-2525 hasbeen deposited with ATCC on Dec. 22, 1998 and is assigned ATCC depositno. 203549.

[0866] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 10 (SEQ ID NO:18), evidenced significant homologybetween the PRO1890 amino acid sequence and the following Dayhoffsequences: AF093673_(—)1, P_W44118, AB014609_(—)1, AC005254_(—)1,AF026547_(—)1, LEC2_MEGRO, PGCV_HUMAN, GEN12667, P_R06331 andCELF52E1_(—)9.

Example 9 Isolation of cDNA clones Encoding Human PRO1887

[0867] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA43041”. Based on this consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO1887.

[0868] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-GCAAAGCTCTGCCTCCTTGGCC-3′; and (SEQ ID NO:24) reverse PCRprimers: 5′-GGGTGGACTGTGCTCTAATGGACGC-3′, and (SEQ ID NO:25)5′-CGTGGCACTGGGTTGATC-3′. (SEQ ID NO:26)

[0869] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA43041 sequence which had the followingnucleotide sequence:

[0870] hybridization probe:5′-GATGCAGTTCTGGTCAGAGACGCTCCCCAGCAAGATACAACAGTG-3′ (SEQ ID NO:27).

[0871] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1887 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human bone marrow.

[0872] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1887, designated herein as“DNA79862-2522” (FIG. 11; SEQ ID NO:22), and the derived proteinsequence for PRO1887.

[0873] DNA79862-2522 is shown in FIG. 11 (SEQ ID NO:22). CloneDNA79862-2522 contains a single open reading frame with an apparenttranslational initiation site at nucleotide positions 6-8, and anapparent stop codon at nucleotide positions 1719-1721. The predictedpolypeptide precursor is 571 amino acids long. The full-length PRO1887protein shown in FIG. 12 has an estimated molecular weight of about62,282 daltons and a pI of about 5.56. Additional features of thePRO1887 protein include a signal peptide at about amino acids 1-27; atransmembrane domain at about amino acids 226-245; a potentialN-glycosylation site at about amino acids 105-108; N-myristoylationsites at about amino acids 10-15, 49-54, 62-67, 86-91, 150-155, 155-160,162-167, 217-222, 227-232, 228-233, 232-237, 262-267, 257-362, and461-466; a prokaryotic membrane lipoprotein lipid attachment site atabout amino acids 12-22; and a carboxylesterases type-B serine activesite at about amino acids 216-231.

[0874] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 12 (SEQ ID NO:23), revealed significant homologybetween the PRO1887 amino acid sequence and Dayhoff sequence ESTM_MOUSE.Homology was also found between the PRO1887 amino acid sequence and thefollowing additional Dayhoff sequences: D50579_(—)1, I61085, EST1_HUMAN,GEN12405, P_W39078, GEN13248, P_R58980, A31800_(—)1, and P_R45189.

[0875] Clone DNA79862-2522 was deposited with the ATCC on Dec. 22, 1998,and is assigned ATCC deposit no. 203550.

Example 10 Isolation of cDNA clones Encoding Human PRO1785

[0876] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA35718”. Based on the DNA35718consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO1785.

[0877] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-ATCCTCCAACATGGAGCCTCTTGC-3′; (SEQ ID NO:30) forward PCRprimer: 5′-GTATCTTGTCAACCCTGAGG-3′; and (SEQ ID NO:31) reverse PCRprimer: 5′-TAACCAGAGCTGCTATGTCAGGCC-3′; (SEQ ID NO:32)

[0878] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA35718 sequence which had the followingnucleotide sequence:

[0879] hybridization probe:5′-AGGCAAAGTTTCACTAGTTGTAAACGTGGCCAGTGACTGCCAACTCACAG-3′ (SEQ ID NO:33).

[0880] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO1785 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human aorticendothelial cells.

[0881] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO1785 (designated herein as DNA80136-2503[FIG. 13, SEQ ID NO:28]; and the derived protein sequence for PRO1785.

[0882] The entire coding sequence of PRO1785 is shown in FIG. 13 (SEQ IDNO:28). Clone DNA80136-2503 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 2-4 andan apparent stop codon at nucleotide positions 629-631 of SEQ ID NO:28.The predicted polypeptide precursor is 209 amino acids long. There is asignal peptide at about amino acids 1-31, a transmembrane domain atabout amino acids 18-37 and a glutathione peroxidase signature at aboutamino acids 104-111 of SEQ ID NO:29. Clone DNA80136-2503 has beendeposited with the ATCC and is assigned ATCC deposit no. 203541. Thefull-length PRO1785 protein shown in FIG. 14 has an estimated molecularweight of about 23,909 daltons and a pI of about 9.68.

[0883] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a NU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 14 (SEQ ID NO:29), revealed sequence identitybetween the PRO1785 amino acid sequence and the following Dayhoffsequences: GSHC_SCHMA, P_R44988, AB012395_(—)1, GSHH_HUMAN,AC004151_(—)3, BTUE ECOLI, GSHC_HUMAN, P_R89910, PWU88907_(—)1 andD379161.

Example 11 Isolation of cDNA clones Encoding Human PRO4353

[0884] A consensus DNA sequence was assembled relative to other ESTsequences using repeated cycles of BLAST and phrap as described inExample I above. This consensus sequence is designated herein“DNA39482”. Based on the DNA39482 consensus sequence, oligonucleotideswere synthesized: 1) to identify by PCR a cDNA library that containedthe sequence of interest, and 2) for use as probes to isolate a clone ofthe full-length coding sequence for PRO4353.

[0885] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-GAGGACCTACCGGCCGGACAG-3′ and (SEQ ID NO:36) reverse PCRprimer: 5′-ATACACCCCGAGTACTGCTGGCAG-3′. (SEQ ID NO:37)

[0886] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA39482 sequence which had the followingnucleotide sequence:

[0887] hybridization probe:5′-AGACAGGGCAGCGGCTGCTGAGCTTGGAGCTGGACGCAGCTT-3′ (SEQ ID NO:38).

[0888] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO4353 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human aorticendothelial cells.

[0889] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO4353 (designated herein as DNA80145-2594[FIG. 15, SEQ ID NO:34]; and the derived protein sequence for PRO4353.

[0890] The entire coding sequence of PRO4353 is shown in FIG. 15 (SEQ IDNO:34). Clone DNA80145-2594 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 19-21,and an apparent stop codon at nucleotide positions 2683-2685. Thepredicted polypeptide precursor is 888 amino acids long. CloneDNA80145-2594 has been deposited with ATCC and is assigned ATCC depositno. 204-PTA. The full-length PRO4353 protein shown in FIG. 16 has anestimated molecular weight of about 95,285 daltons and a pI of about8.89.

[0891] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 16 (SEQ ID NO:34), revealed homology between thePRO4353 amino acid sequence and the following Dayhoff sequences(sequences and related text incorporated herein): P_W19857,AB000776_(—)1, P_W57260, JH0798, P_R71382, CEY54E5B_(—)1, I48747,MUSC_(—)1, P_R71383 and P_W63748.

Example 12 Isolation of cDNA clones Encoding Human PRO4357

[0892] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA80155”. Based on the DNA80155consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO4357.

[0893] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-GAAGGTGGAAATTAAATTCCAAGGGC-3′ and (SEQ ID NO:41) reverse PCRprimer: 5′-CGATAAGCTGCTACAGTGCCATCG-3′. (SEQ ID NO:42)

[0894] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA80155 sequence which had the followingnucleotide sequence:

[0895] hybridization probe:5′-GTGACTGTCCTCTGCAAGATAGTGCAGCCTGGCTACGGGA-3′ (SEQ ID NO:43).

[0896] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO4357 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human aorticendothelial cells.

[0897] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO4357; and the derived protein sequencefor PRO4357.

[0898] The entire coding sequence of PRO4357 is shown in FIG. 17 (SEQ IDNO:39). Clone DNA84917-2597 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 286-288,and an apparent stop codon at nucleotide positions 1792-1794. Thepredicted polypeptide precursor is 502 amino acids long. CloneDNA84917-2597 has been deposited with ATCC and is assigned ATCC depositno. 203863. The full-length PRO4357 protein shown in FIG. 18 has anestimated molecular weight of about 58,043 daltons and a pi of about7.94.

[0899] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 18 (SEQ ID NO:40), revealed homology between thePRO4357 amino acid sequence and the following Dayhoff sequences:P_W48804, AF003534_(—)66, ATAC00466519, LPSA_BACNO, GELA_DICDI,EHU70560_(—)1, AF089841_(—)1, ABP2_HMAN, P_W19349 and A49551.

Example 13 Isolation of cDNA clones Encoding Human PRO4405

[0900] A consensus DNA sequence was assembled relative to other ESTsequences using repeated cycles of BLAST and phrap. This consensussequence is designated herein “DNA80170”. Based on the DNA80170consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO4405.

[0901] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-CGGGACTTTCGCTACCTGTTGC-3′ and (SEQ ID NO:46) reverse PCRprimer: 5′-CATCATATTCCACAAAATGCTTTGGG-3′. (SEQ ID NO:47)

[0902] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus sequence which had the followingnucleotide sequence:

[0903] hybridization probe: 5′-CCTTCGGGGATTCTTCCCGGCTCCCGTTCGTTCCTCTG-3′(SEQ ID NO:48).

[0904] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO4405 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal kidney.

[0905] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO4405 (designated herein as DNA84920-2614[FIG. 19, SEQ ID NO:44]; and the derived protein sequence for PRO4405.

[0906] The entire coding sequence of PRO4405 is shown in FIG. 19 (SEQ IDNO:44). Clone DNA84920-2614 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 79-81,and an apparent stop codon at nucleotide positions 1009-1011. Thepredicted polypeptide precursor is 310 amino acids long. CloneDNA84920-2614 has been deposited with ATCC and is assigned ATCC depositno. 203966. The full-length PRO4405 protein shown in FIG. 20 has anestimated molecular weight of about 33,875 daltons and a 131 of about7.08.

[0907] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 20 (SEQ ID NO:45), revealed homology between thePRO4405 amino acid sequence and the following Dayhoff sequences(sequences and related text incorporated herein): YA93_SCHPO, S62432,YJG2_YEAST, AC004472_(—)3, AB004539_(—)7, S64782, CELC27A12_(—)8,AF109219_(—)1, AF086791_(—)10, and P_W75859.

Example 14 Isolation of cDNA clones Encoding Human PRO4356

[0908] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA80200”. Based upon an observedhomology between the DNA80200 consensus sequence and an EST sequencecontained within Merck EST clone 248287, Merck EST clone 248287 waspurchased and its insert obtained and sequenced, thereby providingDNA86576-2595.

[0909] The entire coding sequence of PRO4356 is shown in FIG. 21 (SEQ IDNO:49). Clone DNA86576-2595 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 55-57,and an apparent stop codon at nucleotide positions 808-810. Thepredicted polypeptide precursor is 251 amino acids long. CloneDNA86576-2595 has been deposited with ATCC and is assigned ATCC depositno. 203868. The full-length PRO4356 protein shown in FIG. 22 has anestimated molecular weight of about 26,935 daltons and a pI of about7.42.

[0910] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 22 (SEQ ID NO:50), revealed homology between thePRO4356 amino acid sequence and the following Dayhoff sequencesincorporated herein: RNMAGPIAN_(—)1, UPAR_BOVIN, S42152, AF007789_(—)1,UPAR_RAT, UPAR_MOUSE, P_W31165, P_W31168, P_R44423 and P_W26359.

Example 15 Isolation of cDNA clones Encoding Human PRO4352

[0911] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is designated herein “DNA83397”. Based on the DNA83397consensus sequence, oligonucleotides were synthesized: 1) to identify byPCR a cDNA library that contained the sequence of interest, and 2) foruse as probes to isolate a clone of the full-length coding sequence forPRO4352.

[0912] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-CTGGGGAGTGTCCTTGGCAGGTTC-3′ and (SEQ ID NO:53) reverse PCRprimer: 5′-CAGCATACAGGGCTTTAGGGCACAC-3′. (SEQ ID NO:54)

[0913] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA83397 sequence which had the followingnucleotide sequence:

[0914] hybridization probe:5′-CGGTGACTGAGGAAACAGAGAAAGGATCCTTTGTGGTCAATCTGGC-3′ (SEQ ID NO:55).

[0915] In order to screen several libraries for a source of afull-length clone, DNA from the libraries was screened by PCRamplification with the PCR primer pair identified above. A positivelibrary was then used to isolate clones encoding the PRO4352 gene usingthe probe oligonucleotide and one of the PCR primers. RNA forconstruction of the cDNA libraries was isolated from human fetal brain.

[0916] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for PRO4352 (designated herein as DNA87976-2593[FIG. 23, SEQ ID NO:51]; and the derived protein sequence for PRO4352.

[0917] The entire coding sequence of PRO4352 is shown in FIG. 23 (SEQ IDNO:51). Clone DNA87976-2593 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 179-181,and an apparent stop codon at nucleotide positions 2579-2581 of SEQ IDNO:51. The predicted polypeptide precursor is 800 amino acids long.Clone DNA87976-2593 has been deposited with ATCC and is assigned ATCCdeposit no. 203888. The full-length PRO4352 protein shown in FIG. 24 hasan estimated molecular weight of about 87,621 daltons and a p1 of about4.77.

[0918] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 24 (SEQ ID NO:52), revealed homology between thePRO4352 amino acid sequence and the following Dayhoff sequences:P_R86865, P_R86866, RATPCDH_(—)1, AB011160_(—)1, MMU88549_(—)1,D86917_(—)1, AB008179_(—)1, P_R58907,HSHFATPRO_(—)1, and AF031572_(—)1.

Example 16 Isolation of cDNA clones Encoding Human PRO4380

[0919] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA79132. In light ofDNA79132, DNA92234-2602 was identified.

[0920] The full length clone shown in FIG. 25 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 201-203 and ending at the stop codon found atnucleotide positions 1722-1724 (FIG. 25; SEQ ID NO:56). The predictedpolypeptide precursor (FIG. 26, SEQ ID NO:57) is 507 amino acids long.PRO4380 has a calculated molecular weight of approximately 56,692daltons and an estimated pI of approximately 5.22.

[0921] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 26 (SEQ ID NO:57), revealed homology between thePRO4380 amino acid sequence and the following Dayhoff sequences(sequences and related text incorporated herein): CER11H6_(—)1, S56299,D89150_(—)1, G70870, S43914, LM034616_(—)5, LLU78036_(—)1,AF055904_(—)2, P_W79066 and ARGE_ECOLI.

[0922] Clone DNA92234-2602 was deposited with the ATCC and is assignedATCC deposit no. 203948.

Example 17 Isolation of cDNA clones Encoding Human PRO4354

[0923] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster (92909) sequence designatedherein as DNA10195. This EST cluster sequence was then compared to avariety of expressed sequence tag (EST) databases which included publicEST databases (e.g., GenBank) and a proprietary EST DNA database(LIFESEQ®, Incyte Pharmaceuticals, Palo Alto, Calif.) to identifyexisting homologies. The homology search was performed using thecomputer program BLAST or BLAST2 (Altshul et al., Methods in Enzymology266:460480 (1996)). Those comparisons resulting in a BLAST score of 70(or in some cases 90) or greater that did not encode known proteins wereclustered and assembled into a consensus DNA sequence with the program“phrap” (Phil Green, University of Washington, Seattle, Wash.). Theconsensus sequence obtained therefrom is herein designated as DNA56063.In light of DNA56063, DNA92256-2596 was identified.

[0924] The full length clone shown in FIG. 27 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 108-110 and ending at the stop codon found atnucleotide positions 852-854 (FIG. 27; SEQ ID NO:58). The predictedpolypeptide precursor (FIG. 28, SEQ ID NO:59) is 248 amino acids long.PRO4354 has a calculated molecular weight of approximately 28,310daltons and an estimated pI of approximately 4.63.

[0925] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 28 (SEQ ID NO:59), revealed homology between thePRO4354 amino acid sequence and the following Dayhoff sequencesincorporated herein: HGS_RF300, CEVK04G11_(—)2,CEC11H1_(—)7,HSU80744_(—)1, CEF09E8_(—)2, RNAJ29671, DDICOI_(—)1,AB020648_(—)1, P_W33887 and A64319.

[0926] Clone DNA92256-2596 was deposited with the ATCC on Mar. 30, 1999and is assigned ATCC deposit no.203891.

Example 18 Isolation of cDNA clones Encoding Human PRO4408

[0927] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA79298. In light ofDNA79298, DNA92274-2617 was identified and sequenced in full.

[0928] The full length clone shown in FIG. 29 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 89-91 and ending at the stop codon found atnucleotide positions 758-760 (FIG. 29; SEQ ID NO:60). The predictedpolypeptide precursor (FIG. 30, SEQ ID NO:61) is 223 amino acids long.PRO4408 has a calculated molecular weight of approximately 25,402daltons and an estimated pI of approximately 8.14.

[0929] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 30 (SEQ ID NO:61), revealed homology between thePRO4408 amino acid sequence and the following Dayhoff sequences:P_R27897, P_R49942, PBP_RAT, CELF40A3_(—)3, D1ONCVO, PC4214, OV16_ONCVO,P_R27718, GEN10789, and OBA5_DROME.

[0930] Clone DNA92274-2617 was deposited with the ATCC and is assignedATCC deposit no. 203971.

Example 19 Isolation of cDNA clones Encoding Human PRO5737

[0931] An expressed sequence tag (EST) DNA database (LIFESEQ®, IncytePharmaceuticals, Palo Alto, Calif.) was searched with a humaninterleukin-1 receptor antagonist (hIL-1Ra) sequence, and an ESTsequence, designated herein as 1433156 was identified, which, showedhomology with the hIL-1Ra known protein. EST clone 1433156 was purchasedfrom Incyte Pharmaceuticals (Palo Alto, Calif.) and the cDNA insert wasobtained and sequenced in its entirety, giving the DNA92929-2534sequence.

[0932] The entire nucleotide sequence of DNA92929-2534 is shown in FIG.31 (SEQ ID NO:62). Clone DNA92929-2534 contains a single open readingframe with an apparent translational initiation site at nucleotidepositions 96-98 and a stop codon at nucleotide positions 498-500 (FIG.31; SEQ ID NO:62). The predicted polypeptide precursor (hIL-1Ra2) is 134amino acids long. The putative signal sequence extends from amino acidpositions 1-17. Clone DNA92929-2534 was deposited with ATCC and wasassigned ATCC deposit no. 203586. The full-length hIL-lra2 protein shownin FIG. 32 has an estimated molecular weight of about 14,927 daltons anda pI of about 4.8.

[0933] Based on a BLAST and FastA sequence alignment analysis (using theALIGN-2 computer program) of the full-length sequence, hIL-1Ra2 (FIG.32, SEQ ID NO:63) shows significant amino acid sequence identity tohIL-1Rαp protein. hIL-1Ra2 is believed to be a splice variant ofhIL-1Rαβ.

Example 20 Isolation of cDNA clones Encoding Human PRO4425

[0934] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA81099.

[0935] In light of an observed sequence homology between the DNA81099sequence and an EST sequence contained within the EST clone no.AA448744, the EST clone AA448744 was purchased from Merck and the cDNAinsert was obtained and sequenced. The sequence of this cDNA insert isshown in FIG. 33 and is herein designated as DNA93011-2637.

[0936] The full length clone shown in FIG. 33 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 27-29 and ending at the stop codon found atnucleotide positions 435-437 (FIG. 33; SEQ ID NO:64). The predictedpolypeptide precursor (FIG. 34, SEQ ID NO:65) is 136 amino acids long.PRO4425 has a calculated molecular weight of approximately 15,577daltons and an estimated pI of approximately 8.88.

[0937] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 34 (SEQ ID NO:65), revealed homology between thePRO4425 amino acid sequence and the following Dayhoff sequences:HGS_RE295, S44655, YOJ8_CAEEL, VBR1_CLVK, P_R39520, P_R65332, P_R39388,TGL4_HUMAN, YKAB_CAEEL, and S71105.

[0938] Clone DNA93011-2637 was deposited with the ATCC and is assignedATCC deposit no. 20-PTA.

Example 21 Isolation of cDNA clones Encoding Human PRO5990

[0939] A consensus DNA sequence was assembled relative to other ESTsequences using phrap as described in Example 1 above. This consensussequence is herein designated DNA86602. Based on the DNA86602 consensussequence, oligonucleotides were synthesized: 1) to identify by PCR acDNA library that contained the sequence of interest, and 2) for use asprobes to isolate a clone of the full-length coding sequence forPRO5990.

[0940] PCR primers (forward and reverse) were synthesized: forward PCRprimer: 5′-CGTCACAGGAACTTCAGCACCC-3′ (SEQ ID NO:68) reverse PCR primer:5′-GTCTTGGCTTCCTCCAGGTTTGG-3′ (SEQ ID NO:69)

[0941] Additionally, a synthetic oligonucleotide hybridization probe wasconstructed from the consensus DNA86602 sequence which had the followingnucleotide sequence

[0942] hybridization probe:

[0943] 5′-GGACAGCGCTCCCCTCTACCTGGAGACTTGACTCCCGC-3′ (SEQ ID NO:70)

[0944] RNA for construction of the cDNA libraries was isolated fromhuman fetal brain tissue.

[0945] DNA sequencing of the clones isolated as described above gave thefull-length DNA sequence for a full-length PRO5990 polypeptide(designated herein as DNA96042-2682 [FIG. 35, SEQ ID NO:66]) and thederived protein sequence for that PRO5990 polypeptide.

[0946] The full length clone identified above contained a single openreading frame with an apparent translational initiation site atnucleotide positions 265-267 and a stop signal at nucleotide positions1669-1671 (FIG. 35, SEQ ID NO:66). The predicted polypeptide precursoris 468 amino acids long, has a calculated molecular weight ofapproximately 53,005 daltons and an estimated pI of approximately 4.98.Analysis of the full-length PRO5990 sequence shown in FIG. 36 (SEQ IDNO:67) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 36, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA96042-2682 has been deposited with ATCC on Jul. 20, 1999 and isassigned ATCC deposit no. 382-PTA.

[0947] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 36 (SEQ ID NO:67), evidenced sequence identitybetween the PRO5990 amino acid sequence and the following Dayhoffsequences: SG3_MOUSE; SG3_RAT; GEN14673; ENHMHCAX_(—)1; MYS2_DICDI;NFU43192_(—)1; US01_YEAST; A56577; PFLSA13_(—)1; CELF12F3_(—)3.

Example 22 Isolation of cDNA clones Encoding Human PRO6030

[0948] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the LIFESEQ®(Incyte Pharmaceuticals, Palo Alto, Calif.) database, designated hereinas CLU20900. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., Genbank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA81229.

[0949] In light of an observed sequence homology between the DNA81229sequence and an EST sequence encompassed within clone no. 4020130H1 fromthe Incyte (Incyte Pharmaceuticals, Palo Alto, Calif.) database, cloneno.4020130H1 was purchased and the cDNA insert was obtained andsequenced. It was found herein that that cDNA insert encoded afull-length protein. The sequence of this cDNA insert is shown in FIG.37 and is herein designated as DNA96850-2705.

[0950] Clone DNA96850-2705 contains a single open reading frame with anapparent translational initiation site at nucleotide positions 60-62 andending at the stop codon at nucleotide positions 1026-1028 (FIG. 37).The predicted polypeptide precursor is 322 amino acids long (FIG. 38).The full-length PRO6030 protein shown in FIG. 38 has an estimatedmolecular weight of about 34,793 daltons and a pI of about 6.34.Analysis of the full-length PRO6030 sequence shown in FIG. 38 (SEQ IDNO:72) evidences the presence of a variety of important polypeptidedomains as shown in FIG. 38, wherein the locations given for thoseimportant polypeptide domains are approximate as described above. CloneDNA96850-2705 has been deposited with ATCC on Aug. 3, 1999 and isassigned ATCC Deposit No. 479-PTA.

[0951] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using the ALIGN-2 sequence alignment analysis of the full-lengthsequence shown in FIG. 38 (SEQ ID NO:72), evidenced sequence identitybetween the PRO6030 amino acid sequence and the following Dayhoffsequences: AF059571_(—)1; I38346; AF035835_(—)1; P_W83138; P_R54714;P_R65166; P_P93995; BGP1_HUMAN; P_WO6873; A43165_(—)1.

Example 23 Isolation of cDNA clones Encoding Human PRO4424

[0952] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The extendedconsensus sequence obtained therefrom is designated DNA80820. In lightof DNA80820, DNA96857-2636 was identified and sequenced.

[0953] The full length clone shown in FIG. 39 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 52-54 and ending at the stop codon found atnucleotide positions 715-717 (FIG. 39; SEQ ID NO:73). The predictedpolypeptide precursor (FIG. 40, SEQ ID NO:74) is 221 amino acids long.PRO1424 has a calculated molecular weight of approximately 23,598daltons and an estimated pI of approximately 6.96.

[0954] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 40 (SEQ ID NO:74), revealed homology between thePRO4424 amino acid sequence and the following Dayhoff sequences:HGS_A135, JC5105, P_R88555, JC5106, P_R88556, CELR12E2_(—)13, DMC34F38,ATG13D4_(—)7,HGS_A204, S58331.

[0955] Clone DNA96857-2636 was deposited with the ATCC and is assignedATCC deposit no. 17-PTA.

Example 24 Isolation of cDNA clones Encoding Human PRO4422

[0956] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA80134. In light ofDNA80134, DNA96867-2620 was identified and sequenced in full.

[0957] The full length clone shown in FIG. 41 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 318-320 and ending at the stop codon found atnucleotide positions 900-902 (FIG. 41; SEQ ID NO:75). The predictedpolypeptide precursor (FIG. 42, SEQ ID NO:76) is 194 amino acids long.PRO4422 has a calculated molecular weight of approximately 21,431daltons and an estimated pI of approximately 8.57.

[0958] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 42 (SEQ ID NO:76), revealed homology between thePRO4422 amino acid sequence and the following Dayhoff sequences:LYG_CHICK, LYG_CYGAT, LYG_ANSAN, LYG_STRCA, P_W69515, ATAC003680_(—)7,ACCA_HAEIN, I64065, A70853 and AF074611_(—)71.

[0959] Clone DNA96867-2620 was deposited with the ATCC and is assignedATCC deposit no. 203972.

Example 25 Isolation of cDNA clones Encoding Human PRO4430

[0960] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The extendedconsensus sequence obtained therefrom is herein designated DNA82380. Inlight of DNA82380, DNA96878-2626 was identified and sequenced.

[0961] The full length clone shown in FIG. 43 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 56-58 and ending at the stop codon found atnucleotide positions 431-433 (FIG. 43; SEQ ID NO:77). The predictedpolypeptide precursor (FIG. 44, SEQ ID NO:78) is 125 amino acids long.PRO4430 has a calculated molecular weight of approximately 13,821daltons and an estimated pI of approximately 8.6.

[0962] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 44 (SEQ ID NO:78), revealed homology between thePRO4430 amino acid sequence and the following Dayhoff sequences:MMHC213L3_(—)9, A45835, D45835, UPAR_MOUSE, AF043498_(—)1, P_W62066,LY6C_MOUSE, LY6A_MOUSE, P_R58710, and P_R86315.

[0963] Clone DNA96878-2626 was deposited with the ATCC and is assignedATCC deposit no. 23-PTA.

Example 26 Isolation of cDNA clones Encoding Human PRO4499

[0964] Use of the signal sequence algorithm described in Example 3 aboveallowed identification of an EST cluster sequence from the Incytedatabase. This EST cluster sequence was then compared to a variety ofexpressed sequence tag (EST) databases which included public ESTdatabases (e.g., GenBank) and a proprietary EST DNA database (LIFESEQ®,Incyte Pharmaceuticals, Palo Alto, Calif.) to identify existinghomologies. The homology search was performed using the computer programBLAST or BLAST2 (Altshul et al., Methods in Enzymology 266:460-480(1996)). Those comparisons resulting in a BLAST score of 70 (or in somecases 90) or greater that did not encode known proteins were clusteredand assembled into a consensus DNA sequence with the program “phrap”(Phil Green, University of Washington, Seattle, Wash.). The consensussequence obtained therefrom is herein designated DNA81155. In light ofDNA81155, DNA96889-2641 was identified and sequenced.

[0965] The full length clone shown in FIG. 45 contained a single openreading frame with an apparent translational initiation site atnucleotide positions 185-187 and ending at the stop codon found atnucleotide positions 1202-1204 (FIG. 45; SEQ ID NO:79). The predictedpolypeptide precursor (FIG. 46, SEQ ID NO:80) is 339 amino acids long.PRO4499 has a calculated molecular weight of approximately 36,975daltons and an estimated pI of approximately 7.85.

[0966] An analysis of the Dayhoff database (version 35.45 SwissProt 35),using a WU-BLAST2 sequence alignment analysis of the full-lengthsequence shown in FIG. 46 (SEQ ID NO:80), revealed homology between thePRO4499 amino acid sequence and the following Dayhoff sequences:CEF38B7_(—)4, D70575, AF073993_(—)1, PNAPA_(—)1, AF098967_(—)1,AF007140_(—)1, ROA3_HUMAN, E70969, CEY53Cl2B_(—)5 and CEY53C12B_(—)6.

[0967] Clone DNA964389-2641 was deposited with the ATCC and is assignedATCC deposit no. 119-PTA.

Example 27 Use of PRO as a Hybridization Probe

[0968] The following method describes use of a nucleotide sequenceencoding PRO as a hybridization probe.

[0969] DNA comprising the coding sequence of full-length or mature PROas disclosed herein is employed as a probe to screen for homologous DNAs(such as those encoding naturally-occurring variants of PRO) in humantissue cDNA libraries or human tissue genomic libraries.

[0970] Hybridization and washing of filters containing either libraryDNAs is performed under the following high stringency conditions.Hybridization of radiolabeled PRO-derived probe to the filters isperformed in a solution of 50% formamide, 5× SSC, 0.1% SDS, 0.1% sodiumpyrophosphate, 50 mM sodium phosphate, pH 6.8, 2× Denhardt's solution,and 10% dextran sulfate at 42° C. for 20 hours. Washing of the filtersis performed in an aqueous solution of 0.1× SSC and 0.1% SDS at 42° C.

[0971] DNAs having a desired sequence identity with the DNA encodingfull-length native sequence PRO can then be identified using standardtechniques known in the art.

Example 28 Expression of PRO in E. coli

[0972] This example illustrates preparation of an unglycosylated form ofPRO by recombinant expression in E. coli.

[0973] The DNA sequence encoding PRO is initially amplified usingselected PCR primers. The primers should contain restriction enzymesites which correspond to the restriction enzyme sites on the selectedexpression vector. A variety of expression vectors may be employed. Anexample of a suitable vector is pBR322 (derived from E. coli; seeBolivar et al., Gene, 2:95 (1977)) which contains genes for ampicillinand tetracycline resistance. The vector is digested with restrictionenzyme and dephosphorylated. The PCR amplified sequences are thenligated into the vector. The vector will preferably include sequenceswhich encode for an antibiotic resistance gene, a trp promoter, apolyhis leader (including the first six STII codons, polyhis sequence,and enterokinase cleavage site), the PRO coding region, lambdatranscriptional terminator, and an argU gene.

[0974] The ligation mixture is then used to transform a selected E. colistrain using the methods described in Sambrook et al., supra.Transformants are identified by their ability to grow on LB plates andantibiotic resistant colonies are then selected. Plasmid DNA can beisolated and confirmed by restriction analysis and DNA sequencing.

[0975] Selected clones can be grown overnight in liquid culture mediumsuch as LB broth supplemented with antibiotics. The overnight culturemay subsequently be used to inoculate a larger scale culture. The cellsare then grown to a desired optical density, during which the expressionpromoter is turned on.

[0976] After culturing the cells for several more hours, the cells canbe harvested by centrifugation. The cell pellet obtained by thecentrifugation can be solubilized using various agents known in the art,and the solubilized PRO protein can then be purified using a metalchelating column under conditions that allow tight binding of theprotein.

[0977] PRO may be expressed in E. coli in a poly-His tagged form, usingthe following procedure. The DNA encoding PRO is initially amplifiedusing selected PCR primers. The primers will contain restriction enzymesites which correspond to the restriction enzyme sites on the selectedexpression vector, and other useful sequences providing for efficientand reliable translation initiation, rapid purification on a metalchelation column, and proteolytic removal with enterokinase. ThePCR-amplified, poly-His tagged sequences are then ligated into anexpression vector, which is used to transform an E. coli host based onstrain 52 (W3110 fuhA(tonA) lon galE rpoHts(htpRts) clpP(laclq).Transformants are first grown in LB containing 50 mg/ml carbenicillin at30° C. with shaking until an O.D.600 of 3-5 is reached. Cultures arethen diluted 50-100 fold into CRAP media (prepared by mixing 3.57 g(NH₄)₂SO₄, 0.71 g sodium citrate.2H2O, 1.07 g KCl, 5.36 g Difco yeastextract, 5.36 g Sheffield hycase SF in 500 mL water, as well as 110 mMMPOS, pH 7.3, 0.55% (w/v) glucose and 7 mM MgSO₄) and grown forapproximately 20-30 hours at 30° C. with shaking. Samples are removed toverify expression by SDS-PAGE analysis, and the bulk culture iscentrifuged to pellet the cells. Cell pellets are frozen untilpurification and refolding.

[0978]E. coli paste from 0.5 to 1 L fermentations (6-10 g pellets) isresuspended in 10 volumes (w/v) in 7 M guanidine, 20 mnM Tris, pH 8buffer. Solid sodium sulfite and sodium tetrathionate is added to makefinal concentrations of 0.1M and 0.02 M, respectively, and the solutionis stirred overnight at 4° C. This step results in a denatured proteinwith all cysteine residues blocked by sulfitolization. The solution iscentrifuged at 40,000 rpm in a Beckman Ultracentifuge for 30 min. Thesupernatant is diluted with 3-5 volumes of metal chelate column buffer(6 M gulanidine, 20 mM Tris, pH 7.4) and filtered through 0.22 micronfilters to clarify. The clarified extract is loaded onto a 5 ml QiagenNi-NTA metal chelate column equilibrated in the metal chelate columnbuffer. The column is washed with additional buffer containing 50 mMimidazole (Calbiochem, Utrol grade), pH 7.4. The protein is eluted withbuffer containing 250 mM imidazole. Fractions containing the desiredprotein are pooled and stored at 4° C. Protein concentration isestimated by its absorbance at 280 nm using the calculated extinctioncoefficient based on its amino acid sequence.

[0979] The proteins are refolded by diluting the sample slowly intofreshly prepared refolding buffer consisting of: 20 mM Tris, pH 8.6, 0.3M NaCl, 2.5 M urea, 5 mM cysteine, 20 mM glycine and 1 mM EDTA.Refolding volumes are chosen so that the final protein concentration isbetween 50 to 100 micrograms/ml. The refolding solution is stirredgently at 4° C. for 12-36 hours. The refolding reaction is quenched bythe addition of TFA to a final concentration of 0.4% (pH ofapproximately 3). Before further purification of the protein, thesolution is filtered through a 0.22 micron filter and acetonitrile isadded to 2-10% final concentration. The refolded protein ischromatographed on a Poros R1/H reversed phase column using a mobilebuffer of 0.1% TFA with elution with a gradient of acetonitrile from 10to 80%. Aliquots of fractions with A280 absorbance are analyzed on SDSpolyacrylamide gels and fractions containing homogeneous refoldedprotein are pooled. Generally, the properly refolded species of mostproteins are eluted at the lowest concentrations of acetonitrile sincethose species are the most compact with their hydrophobic interiorsshielded from interaction with the reversed phase resin. Aggregatedspecies are usually eluted at higher acetonitrile concentrations. Inaddition to resolving misfolded forms of proteins from the desired form,the reversed phase step also removes endotoxin from the samples.

[0980] Fractions containing the desired folded PRO polypeptide arepooled and the acetonitrile removed using a gentle stream of nitrogendirected at the solution. Proteins are formulated into 20 mM Hepes, pH6.8 with 0.14 M sodium chloride and 4% mannitol by dialysis or by gelfiltration using G25 Superfine (Pharmacia) resins equilibrated in theformulation buffer and sterile filtered.

[0981] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 29 Expression of PRO in Mammalian Cells

[0982] This example illustrates preparation of a potentiallyglycosylated form of PRO by recombinant expression in mammalian cells.

[0983] The vector, pRK5 (see EP 307,247, published Mar. 15, 1989), isemployed as the expression vector. Optionally, the PRO DNA is ligatedinto pRK5 with selected restriction enzymes to allow insertion of thePRO DNA using ligation methods such as described in Sambrook et al.,supra. The resulting vector is called pRK5-PRO.

[0984] In one embodiment, the selected host cells may be 293 cells.Human 293 cells (ATCC CCL 1573) are grown to confluence in tissueculture plates in medium such as DMEM supplemented with fetal calf serumand optionally, nutrient components and/or antibiotics. About 10 μgpRK5-PRO DNA is mixed with about 1 μg DNA encoding the VA RNA gene[Thimmappaya et al., Cell, 31:543 (1982)] and dissolved in 500 μl of 1mM Tris-HCl, 0.1 mM EDTA, 0.227 M CaCl₂. To this mixture is added,dropwise, 500 μl of 50 mM HEPES (pH 7.35), 280 mM NaCl, 1.5 mM NaPO₄,and a precipitate is allowed to form for 10 minutes at 25° C. Theprecipitate is suspended and added to the 293 cells and allowed tosettle for about four hours at 37° C. The culture medium is aspiratedoff and 2 ml of 20% glycerol in PBS is added for 30 seconds. The 293cells are then washed with serum free medium, fresh medium is added andthe cells are incubated for about 5 days.

[0985] Approximately 24 hours after the transfections, the culturemedium is removed and replaced with culture medium (alone) or culturemedium containing 200 μCi/ml ³⁵S-cysteine and 200 μCi/ml ³⁵S-methionine.After a 12 hour incubation, the conditioned medium is collected,concentrated on a spin filter, and loaded onto a 15% SDS gel. Theprocessed gel may be dried and exposed to film for a selected period oftime to reveal the presence of PRO polypeptide. The cultures containingtransfected cells may undergo further incubation (in serum free medium)and the medium is tested in selected bioassays.

[0986] In an alternative technique, PRO may be introduced into 293 cellstransiently using the dextran sulfate method described by Somparyrac etal., Proc. Natl. Acad. Sci., 12:7575 (1981). 293 cells are grown tomaximal density in a spinner flask and 700 μg pRK5-PRO DNA is added. Thecells are first concentrated from the spinner flask by centrifugationand washed with PBS. The DNA-dextran precipitate is incubated on thecell pellet for four hours. The cells are treated with 20% glycerol for90 seconds, washed with tissue culture medium, and re-introduced intothe spinner flask containing tissue culture medium, 5 μg/ml bovineinsulin and 0.1 μg/ml bovine transferrin. After about four days, theconditioned media is centrifuged and filtered to remove cells anddebris. The sample containing expressed PRO can then be concentrated andpurified by any selected method, such as dialysis and/or columnchromatography.

[0987] In another embodiment, PRO can be expressed in CHO cells. ThepRK5-PRO can be transfected into CHO cells using known reagents such asCaPO₄ or DEAE-dextran. As described above, the cell cultures can beincubated, and the medium replaced with culture medium (alone) or mediumcontaining a radiolabel such as ³⁵S-methionine. After determining thepresence of PRO polypeptide, the culture medium may be replaced withserum free medium. Preferably, the cultures are incubated for about 6days, and then the conditioned medium is harvested. The mediumcontaining the expressed PRO can then be concentrated and purified byany selected method.

[0988] Epitope-tagged PRO may also be expressed in host CHO cells. ThePRO may be subcloned out of the pRK5 vector. The subclone insert canundergo PCR to fuse in frame with a selected epitope tag such as apoly-his tag into a Baculovirus expression vector. The poly-his taggedPRO insert can then be subcloned into a SV40 driven vector containing aselection marker such as DHFR for selection of stable clones. Finally,the CHO cells can be transfected (as described above) with the SV40driven vector. Labeling may be performed, as described above, to verifyexpression. The culture medium containing the expressed poly-His taggedPRO can then be concentrated and purified by any selected method, suchas by Ni²⁺-chelate affinity chromatography.

[0989] PRO may also be expressed in CHO and/or COS cells by a transientexpression procedure or in CHO cells by another stable expressionprocedure.

[0990] Stable expression in CHO cells is performed using the followingprocedure. The proteins are expressed as an IgG construct(inimunoadhesin), in which the coding sequences for the soluble forms(e.g. extracellular domains) of the respective proteins are fused to anIgGl constant region sequence containing the hinge, CH2 and CH2 domainsand/or is a poly-His tagged form.

[0991] Following PCR amplification, the respective DNAs are subcloned ina CHO expression vector using standard techniques as described inAusubel et al., Current Protocols of Molecular Biology, Unit 3.16, JohnWiley and Sons (1997). CHO expression vectors are constructed to havecompatible restriction sites 5′ and 3′ of the DNA of interest to allowthe convenient shuttling of cDNA's. The vector used expression in CHOcells is as described in Lucas et al., Nucl. Acids Res. 24:9 (1774-1779(1996), and uses the SV40 early promoter/enhancer to drive expression ofthe cDNA of interest and dihydrofolate reductase (DHFR). DHFR expressionpermits selection for stable maintenance of the plasmid followingtransfection.

[0992] Twelve micrograms of the desired plasmid DNA is introduced intoapproximately 10 million CHO cells using commercially availabletransfection reagents Superfect® (Quiagen), Dosper® or Fugene®(Boehringer Mannheim). The cells are grown as described in Lucas et al.,supra. Approximately 3×10⁻⁷ cells are frozen in an ampule for furthergrowth and production as described below.

[0993] The ampules containing the plasmid DNA are thawed by placementinto water bath and mixed by vortexing. The contents are pipetted into acentrifuge tube containing 10 mLs of media and centrifuged at 1000 rpmfor 5 minutes. The supernatant is aspirated and the cells areresuspended in 10 mL of selective media (0.2 μm filtered PS20 with 5%0.2 μm diafiltered fetal bovine serum). The cells are then aliquotedinto a 100 mL spinner containing 90 mL of selective media. After 1-2days, the cells are transferred into a 250 mL spinner filled with 150 mLselective growth medium and incubated at 37° C. After another 2-3 days,250 mL, 500 mL and 2000 mL spinners are seeded with 3×10⁵ cells/mL. Thecell media is exchanged with fresh media by centrifugation andresuspension in production medium. Although any suitable CHO media maybe employed, a production medium described in U.S. Pat. No. 5,122,469,issued Jun. 16, 1992 may actually be used. A 3L production spinner isseeded at 1.2×10⁶ cells/mL. On day 0, the cell number pH ie determined.On day 1, the spinner is sampled and sparging with filtered air iscommenced. On day 2, the spinner is sampled, the temperature shifted to33° C., and 30 mL of 500 g/L glucose and 0.6 mL of 10% antifoam (e.g.,35% polydimethylsiloxane emulsion, Dow Corning 365 Medical GradeEmulsion) taken. Throughout the production, the pH is adjusted asnecessary to keep it at around 7.2. After 10 days, or until theviability dropped below 70%, the cell culture is harvested bycentrifugation and filtering through a 0.22 μm filter. The filtrate waseither stored at 4° C. or immediately loaded onto columns forpurification.

[0994] For the poly-His tagged constructs, the proteins are purifiedusing a Ni-NTA column (Qiagen). Before purification, imidazole is addedto the conditioned media to a concentration of 5 mM. The conditionedmedia is pumped onto a 6 ml Ni-NTA column equilibrated in 20 mM Hepes,pH 7.4, buffer containing 0.3 M NaCl and 5 mM imidazole at a flow rateof 4-5 ml/min. at 4° C. After loading, the column is washed withadditional equilibration buffer and the protein eluted withequilibration buffer containing 0.25 M imidazole. The highly purifiedprotein is subsequently desalted into a storage buffer containing 10 mMHepes, 0.14 M NaCl and 4% mannitol, pH 6.8, with a 25 ml G25 Superfine(Pharmacia) column and stored at −80° C.

[0995] Immunoadhesin (Fc-containing) constructs are purified from theconditioned media as follows. The conditioned medium is pumped onto a 5ml Protein A column (Pharmacia) which had been equilibrated in 20 mM Naphosphate buffer, pH 6.8. After loading, the column is washedextensively with equilibration buffer before elution with 100 mM citricacid, pH 3.5. The eluted protein is immediately neutralized bycollecting 1 ml fractions into tubes containing 275 μL of 1 M Trisbuffer, pH 9. The highly purified protein is subsequently desalted intostorage buffer as described above for the poly-His tagged proteins. Thehomogeneity is assessed by SDS polyacrylamide gels and by N-terminalamino acid sequencing by Edman degradation.

[0996] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 30 Expression of PRO in Yeast

[0997] The following method describes recombinant expression of PRO inyeast.

[0998] First, yeast expression vectors are constructed for intracellularproduction or secretion of PRO from the ADH2/GAPDH promoter. DNAencoding PRO and the promoter is inserted into suitable restrictionenzyme sites in the selected plasmid to direct intracellular expressionof PRO. For secretion, DNA encoding PRO can be cloned into the selectedplasmid, together with DNA encoding the ADH2/GAPDH promoter, a nativePRO signal peptide or other mammalian signal peptide, or, for example, ayeast alpha-factor or invertase secretory signal/leader sequence, andlinker sequences (if needed) for expression of PRO.

[0999] Yeast cells, such as yeast strain AB110, can then be transformedwith the expression plasmids described above and cultured in selectedfermentation media. The transformed yeast supernatants can be analyzedby precipitation with 10% trichloroacetic acid and separation bySDS-PAGE, followed by staining of the gels with Coomassie Blue stain.

[1000] Recombinant PRO can subsequently be isolated and purified byremoving the yeast cells from the fermentation medium by centrifugationand then concentrating the medium using selected cartridge filters. Theconcentrate containing PRO may further be purified using selected columnchromatography resins.

[1001] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 31 Expression of PRO in Baculovirus-Infected Insect Cells

[1002] The following method describes recombinant expression of PRO inBaculovirus-infected insect cells.

[1003] The sequence coding for PRO is fused upstream of an epitope tagcontained within a baculovirus expression vector. Such epitope tagsinclude poly-his tags and immunoglobulin tags (like Fc regions of IgG).A variety of plasmids may be employed, including plasmids derived fromcommercially available plasmids such as pVL1393 (Novagen). Briefly, thesequence encoding PRO or the desired portion of the coding sequence ofPRO such as the sequence encoding the extracellular domain of atransmembrane protein or the sequence encoding the mature protein if theprotein is extracellular is amplified by PCR with primers complementaryto the 5′ and 3′ regions. The 5′ primer may incorporate flanking(selected) restriction enzyme sites. The product is then digested withthose selected restriction enzymes and subcloned into the expressionvector.

[1004] Recombinant baculovirus is generated by co-transfecting the aboveplasmid and BaculoGold™ virus DNA (Pharmiingen) into Spodopterafrugiperda (“Sf9”) cells (ATCC CRL 1711) using lipofectin (commerciallyavailable from GIBCO-BRL). After 4-5 days of incubation at 28° C., thereleased viruses are harvested and used for further amplifications.Viral infection and protein expression are performed as described byO'Reilley et al., Baculovirus expression vectors: A Laboratory Manual,Oxford: Oxford University Press (1994).

[1005] Expressed poly-his tagged PRO can then be purified, for example,by Ni²⁺-chelate affinity chromatography as follows. Extracts areprepared from recombinant virus-infected Sf9 cells as described byRupert et al., Nature, 362:175-179 (1993). Briefly, Sf9 cells arewashed, resuspended in sonication buffer (25 mL Hepes, pH 7.9; 12.5 mMMgCl₂; 0.1 mM EDTA; 10% glycerol; 0.1% NP-40; 0.4 M KCI), and sonicatedtwice for 20 seconds on ice. The sonicates are cleared bycentrifugation, and the supernatant is diluted 50-fold in loading buffer(50 mM phosphate, 300 mM NaCl, 10% glycerol, pH 7.8) and filteredthrough a 0.45 μm filter. A Ni²⁺-NTA agarose column (commerciallyavailable from Qiagen) is prepared with a bed volume of 5 mL, washedwith 25 mL of water and equilibrated with 25 mL of loading buffer. Thefiltered cell extract is loaded onto the column at 0.5 mL per minute.The column is washed to baseline A₂₈₀ with loading buffer, at whichpoint fraction collection is started. Next, the column is washed with asecondary wash buffer (50 mM phosphate; 300 mM NaCl, 10% glycerol, pH6.0), which elutes nonspecifically bound protein. After reaching A₂₈₀baseline again, the column is developed with a 0 to 500 mM Imidazolegradient in the secondary wash buffer. One mL fractions are collectedand analyzed by SDS-PAGE and silver staining or Western blot withNi²⁺-NTA-conjugated to alkaline phosphatase (Qiagen). Fractionscontaining the eluted His₁₀-tagged PRO are pooled and dialyzed againstloading buffer.

[1006] Alternatively, purification of the IgG tagged (or Fc tagged) PROcan be performed using known chromatography techniques, including forinstance, Protein A or protein G column chromatography.

[1007] Many of the PRO polypeptides disclosed herein were successfullyexpressed as described above.

Example 32 Preparation of Antibodies that Bind PRO

[1008] This example illustrates preparation of monoclonal antibodieswhich can specifically bind PRO.

[1009] Techniques for producing the monoclonal antibodies are known inthe art and are described, for instance, in Goding, supra. Immunogensthat may be employed include purified PRO, fusion proteins containingPRO, and cells expressing recombinant PRO on the cell surface. Selectionof the immunogen can be made by the skilled artisan without undueexperimentation.

[1010] Mice, such as Balb/c, are immunized with the PRO immunogenemulsified in complete Freund's adjuvant and injected subcutaneously orintraperitoneally in an amount from 1-100 micrograms. Alternatively, theimmunogen is emulsified in MPL-TDM adjuvant (Ribi ImmunochemicalResearch, Hamilton, Mont.) and injected into the animal's hind footpads. The immunized mice are then boosted 10 to 12 days later withadditional immunogen emulsified in the selected adjuvant. Thereafter,for several weeks, the mice may also be boosted with additionalimmunization injections. Serum samples may be periodically obtained fromthe mice by retro-orbital bleeding for testing in ELISA assays to detectanti-PRO antibodies.

[1011] After a suitable antibody titer has been detected, the animals“positive” for antibodies can be injected with a final intravenousinjection of PRO. Three to four days later, the mice are sacrificed andthe spleen cells are harvested. The spleen cells are then fused (using35% polyethylene glycol) to a selected murine myeloma cell line such asP3X63AgU.1, available from ATCC, No. CRL 1597. The fusions generatehybridoma cells which can then be plated in 96 well tissue cultureplates containing HAT (hypoxanthine, aminopterin, and thymidine) mediumto inhibit proliferation of non-fused cells, myeloma hybrids, and spleencell hybrids.

[1012] The hybridoma cells will be screened in an ELISA for reactivityagainst PRO. Determination of “positive” hybridoma cells secreting thedesired monoclonal antibodies against PRO is within the skill in theart.

[1013] The positive hybridoma cells can be injected intraperitoneallyinto syngeneic Balb/c mice to produce ascites containing the anti-PROmonoclonal antibodies. Alternatively, the hybridoma cells can be grownin tissue culture flasks or roller bottles. Purification of themonoclonal antibodies produced in the ascites can be accomplished usingammonium sulfate precipitation, followed by gel exclusionchromatography. Alternatively, affinity chromatography based uponbinding of antibody to protein A or protein G can be employed.

Example 33 Purification of PRO Polypeptides Using Specific Antibodies

[1014] Native or recombinant PRO polypeptides may be purified by avariety of standard techniques in the art of protein purification. Forexample, pro-PRO polypeptide, mature PRO polypeptide, or pre-PROpolypeptide is purified by immunoaffinity chromatography usingantibodies specific for the PRO polypeptide of interest. In general, animmunoaffinity column is constructed by covalently coupling the anti-PROpolypeptide antibody to an activated chromatographic resin.

[1015] Polyclonal immunoglobulins are prepared from immune sera eitherby precipitation with ammonium sulfate or by purification on immobilizedProtein A (Pharmacia LKB Biotechnology, Piscataway, N.J.). Likewise,monoclonal antibodies are prepared from mouse ascites fluid by ammoniumsulfate precipitation or chromatography on immobilized Protein A.Partially purified immunoglobulin is covalently attached to achromatographic resin such as CnBr-activated SEPHAROSE™ (Pharnacia LKBBiotechnology). The antibody is coupled to the resin, the resin isblocked, and the derivative resin is washed according to themanufacturer's instructions.

[1016] Such an imrmunoaffinity column is utilized in the purification ofPRO polypeptide by preparing a fraction from cells containing PROpolypeptide in a soluble form. This preparation is derived bysolubilization of the whole cell or of a subcellular fraction obtainedvia differential centrifugation by the addition of detergent or by othermethods well known in the art. Alternatively, soluble PRO polypeptidecontaining a signal sequence may be secreted in useful quantity into themedium in which the cells are grown.

[1017] A soluble PRO polypeptide-containing preparation is passed overthe immunoaffinity column, and the column is washed under conditionsthat allow the preferential absorbance of PRO polypeptide (e.g., highionic strength buffers in the presence of detergent). Then, the columnis eluted under conditions that disrupt antibody/PRO polypeptide binding(e.g., a low pH buffer such as approximately pH 2-3, or a highconcentration of a chaotrope such as urea or thiocyanate ion), and PROpolypeptide is collected.

Example 34 Drug Screening

[1018] This invention is particularly useful for screening compounds byusing PRO polypeptides or binding fragment thereof in any of a varietyof drug screening techniques. The PRO polypeptide or fragment employedin such a test may either be free in solution, affixed to a solidsupport, borne on a cell surface, or located intracellularly. One methodof drug screening utilizes eukaryotic or prokaryotic host cells whichare stably transformed with recombinant nucleic acids expressing the PROpolypeptide or fragment. Drugs are screened against such transformedcells in competitive binding assays. Such cells, either in viable orfixed form, can be used for standard binding assays. One may measure,for example, the formation of complexes between PRO polypeptide or afragraent and the agent being tested. Alternatively, one can examine thediminution in complex formation between the PRO polypeptide and itstarget cell or target receptors caused by the agent being tested.

[1019] Thus, the present invention provides methods of screening fordrugs or any other agents which can affect a PRO polypeptide-associateddisease or disorder. These methods comprise contacting such an agentwith an PRO polypeptide or fragment thereof and assaying (I) for thepresence of a complex between the agent and the PRO polypeptide orfragment, or (ii) for the presence of a complex between the PROpolypeptide or fragment and the cell, by methods well known in the art.In such competitive binding assays, the PRO polypeptide or fragment istypically labeled. After suitable incubation, free PRO polypeptide orfragment is separated from that present in bound form, and the amount offree or uncomplexed label is a measure of the ability of the particularagent to bind to PRO polypeptide or to interfere with the PROpolypeptide/cell complex.

[1020] Another technique for drug screening provides high throughputscreening for compounds having suitable binding affinity to apolypeptide and is described in detail in WO 84103564, published on Sep.13, 1984. Briefly stated, large numbers of different small peptide testcompounds are synthesized on a solid substrate, such as plastic pins orsome other surface. As applied to a PRO polypeptide, the peptide testcompounds are reacted with PRO polypeptide and washed. Bound PROpolypeptide is detected by methods well known in the art. Purified PROpolypeptide can also be coated directly onto plates for use in theaforementioned drug screening techniques. In addition, non-neutralizingantibodies can be used to capture the peptide and immobilize it on thesolid support.

[1021] This invention also contemplates the use of competitive drugscreening assays in which neutralizing antibodies capable of binding PROpolypeptide specifically compete with a test compound for binding to PROpolypeptide or fragments thereof. In this manner, the antibodies can beused to detect the presence of any peptide which shares one or moreantigenic determinants with PRO polypeptide.

Example 35 Rational Drug Design

[1022] The goal of rational drug design is to produce structural analogsof biologically active polypeptide of interest (i.e., a PRO polypeptide)or of small molecules with which they interact, e.g., agonists,antagonists, or inhibitors. Any of these examples can be used to fashiondrugs which are more active or stable forms of the PRO polypeptide orwhich enhance or interfere with the function of the PRO polypeptide invivo (c.f., Hodgson, Bio/Technology, 9: 19-21 (1991)).

[1023] In one approach, the three-dimensional structure of the PROpolypeptide, or of an PRO polypeptide-inhibitor complex, is determinedby x-ray crystallography, by computer modeling or, most typically, by acombination of the two approaches. Both the shape and charges of the PROpolypeptide must be ascertained to elucidate the structure and todetermine active site(s) of the molecule. Less often, usefullinformation regarding the structure of the PRO polypeptide may be gainedby modeling based on the structure of homologous proteins.

[1024] In both cases, relevant structural information is used to designanalogous PRO polypeptide-like molecules or to identify efficientinhibitors. Useful examples of rational drug design may includemolecules which have improved activity or stability as shown by Braxtonand Wells, Biochemistry. 31:7796-7801 (1992) or which act as inhibitors,agonists, or antagonists of native peptides as shown by Athauda et al.,J. Biochem., 113:742-746 (1993).

[1025] It is also possible to isolate a target-specific antibody,selected by functional assay, as described above, and then to solve itscrystal structure. This approach, in principle, yields a pharmacore uponwhich subsequent drug design can be based. It is possible to bypassprotein crystallography altogether by generating anti-idiotypicantibodies (anti-ids) to a functional, pharmacologically activeantibody. As a mirror image of a mirror image, the binding site of theanti-ids would be expected to be an analog of the original receptor. Theanti-id could then be used to identify and isolate peptides from banksof chemically or biologically produced peptides. The isolated peptideswould then act as the pharmacore.

[1026] By virtue of the present invention, sufficient amounts of the PROpolypeptide may be made available to perform such analytical studies asX-ray crystallography. In addition, knowledge of the PRO polypeptideamino acid sequence provided herein will provide guidance to thoseemploying computer modeling techniques in place of or in addition tox-ray crystallography.

Example 36 Chondrocyte Re-differentiation Assay (Assay 110)

[1027] This assay shows that certain polypeptides of the invention actto induce redifferentiation of chondrocytes, therefore, are expected tobe useful for the treatment of various bone and/or cartilage disorderssuch as, for example, sports injuries and arthritis. The assay isperformed as follows. Porcine chondrocytes are isolated by overnightcollagenase digestion of articulary cartilage ofmetacarpophalangealjoints of 4-6 month old female pigs. The isolatedcells are then seeded at 25,000 cells/cm² in Ham F-12 containing 10% FBSand 4 μg/ml gentamycin. The culture media is changed every third day andthe cells are then seeded in 96 well plates at 5,000 cells/well in 100μl of the same media without serum and 100 μl of the test PROpolypeptide, 5 nM staurosporin (positive control) or medium alone(negative control) is added to give a final volume of 200 μl/well. After5 days of incubation at 37° C., a picture of each well is taken and thedifferentiation state of the chondrocytes is determined. A positiveresult in the assay occurs when the redifferentiation of thechondrocytes is determined to be more similar to the positive controlthan the negative control.

[1028] The following polypeptide tested positive in this assay: PRO1484,PRO1890, PRO1887, PRO4353, PRO4357, PRO4405, PRO5737 and PRO5990.

Example 37 Detection of Polypeptides That Affect Glucose or FFA Uptakein Skeletal Muscle (Assay 106)

[1029] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by skeletal muscle cells.PRO polypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by skeletal muscle would bebeneficial including, for example, diabetes or hyper- orhypo-insulinemia.

[1030] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat differentiated skeletal muscle, and allowed to incubateovernight. Then fresh media with the PRO polypeptide and +/− insulin areadded to the wells. The sample media is then monitored to determineglucose and FFA uptake by the skeletal muscle cells. The insulin willstimulate glucose and FFA uptake by the skeletal muscle, and insulin inmedia without the PRO polypeptide is used as a positive control, and alimit for scoring. As the PRO polypeptide being tested may eitherstimulate or inhibit glucose and FFA uptake, results are scored aspositive in the assay if greater than 1.5 times or less than 0.5 timesthe insulin control.

[1031] The following PRO polypeptides tested positive as eitherstimulators or inhibitors of glucose and/or FFA uptake in this assay:]PRO1484, PRO1122, PRO1889, PRO4357 and PRO4380.

Example 38 Detection of PRO Polypeptides That Affect Glucose or FFAUptake by Primary Rat Adipocytes (Assay 94)

[1032] This assay is designed to determine whether PRO polypeptides showthe ability to affect glucose or FFA uptake by adipocyte cells. PROpolypeptides testing positive in this assay would be expected to beuseful for the therapeutic treatment of disorders where either thestimulation or inhibition of glucose uptake by adipocytes would bebeneficial including, for example, obesity, diabetes or hyper- orhypo-insulinemia.

[1033] In a 96 well format, PRO polypeptides to be assayed are added toprimary rat adipocytes, and allowed to incubate overnight. Samples aretaken at 4 and 16 hours and assayed for glycerol, glucose and FFAuptake. After the 16 hour incubation, insulin is added to the media andallowed to incubate for 4 hours. At this time, a sample is taken andglycerol, glucose and FFA uptake is measured. Media containing insulinwithout the PRO polypeptide is used as a positive reference control. Asthe PRO polypeptide being tested may either stimulate or inhibit glucoseand FFA uptake, results are scored as positive in the assay if greaterthan 1.5 times or less than 0.5 times the insulin control.

[1034] The following PRO polypeptides tested positive as stimulators ofglucose and/or FFA uptake in this assay: PRO1890, PRO1785 and PRO4422.

[1035] The following PRO polypeptides tested positive as inhibitors ofglucose and/or FFA uptake in this assay: PRO4334, PRO4425, PRO4424 andPRO4430.

Example 39 Induction of Pancreatic β-Cell Precursor Differentiation(Assay 89)

[1036] This assay shows that certain polypeptides of the invention actto induce differentiation of pancreatic β-cell precursor cells intomature pancreatic β-cells and, therefore, are useful for treatingvarious insulin deficient states in mammals, including diabetesmellitus. The assay is performed as follows. The assay uses a primaryculture of mouse fetal pancreatic cells and the primary readout is analteration in the expression of markers that represent either β-cellprecursors or mature β-cells. Marker expression is measured by real timequantitative PCR (RTQ-PCR); wherein the marker being evaluated isinsulin.

[1037] The pancreata are dissected from E14 embryos (CD1 mice). Thepancreata are then digested with collagenase/dispase in F12/DMEM at 37°C. for 40 to 60 minutes (collagenase/dispase, 1.37 mg/nil, BoehringerMannheim, #1097113). The digestion is then neutralized with an equalvolume of 5% BSA and the cells are washed once with RPMI1640. At day 1,the cells are seeded into 12-well tissue culture plates (pre-coated withlaminin, 20μg/ml in PBS, Boehringer Mannheim, #124317). Cells frompancreata from 1-2 embryos are distributed per well. The culture mediumfor this primary cuture is 14F/1640. At day 2, the media is removed andthe attached cells washed with RPMI/1640. Two mls of minimal media areadded in addition to the protein to be tested. At day 4, the media isremoved and RNA prepared from the cells and marker expression analyzedby real time quantitative RT-PCR. A protein is considered to be activein the assay if it increases the expression of the relevant β-cellmarker as compared to untreated controls.

[1038] 14F/1640 is RPMI1640 (Gibco) plus the following:

[1039] group A 1:1000

[1040] group B 1:1000

[1041] recombinant human insulin 10 μg/ml

[1042] Aprotinin (50 μg/ml) 1:2000 (Boehringer manheim #981532)

[1043] Bovine pituitary extract (BPE) 60 μg/ml

[1044] Gentamycin 100 ng/ml

[1045] Group A: (in 10 ml PBS)

[1046] Transferrin, 100 mg (Sigma T2252)

[1047] Epidermal Growth Factor, 100 μg (BRL 100004)

[1048] Triiodothyronine, 10 μl of 5×10⁻⁶ M (Sigma T5516)

[1049] Ethanolamine, 100 μl of 10⁻¹ M (Sigma E0135)

[1050] Phosphoethalamine, 100 μl of 10⁻¹ M (Sigma P0503)

[1051] Selenium, 4μL of 10⁻¹ M (Aesar #12574)

[1052] Group C: (in 10 ml 100% ethanol)

[1053] Hydrocortisone, 2 μl of 5×10⁻³ M (Sigma #H0135)

[1054] Progesterone, 100 μl of 1×10⁻³ M (Sigma #P6149)

[1055] Forskolin, 500 μl of 20 mM (Calbiochem #44270)

[1056] Minimal media:

[1057] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml), aprotinin(50 μg/ml) and BPE (15 μg/ml).

[1058] Defined media:

[1059] RPMI 1640 plus transferrin (10 μg/ml), insulin (1 μg/ml),gentamycin (100 ng/ml) and aprotinin (50 μg/ml).

[1060] The following polypeptides were positive in this assay: PRO4356.

Example 40 Fetal lemoglobin Induction in an Erythroblastic Cell Line(Assay 107)

[1061] This assay is useful for screening PRO polypeptides for theability to induce the switch from adult hemoglobin to fetal hemoglobinin an erythroblastic cell line. Molecules testing positive in this assayare expected to be useful for therapeutically treating various mammalianhemoglobin-associated disorders such as the various thalassemias. Theassay is performed as follows. Erythroblastic cells are plated instandard growth medium at 1000 cells/well in a 96 well format. PROpolypeptides are added to the growth medium at a concentration of 0.2%or 2% and the cells are incubated for 5 days at 37° C. As a positivecontrol, cells are treated with 100 μM hemin and as a negative control,the cells are untreated. After 5 days, cell lysates are prepared andanalyzed for the expression of gamma globin (a fetal marker). A positivein the assay is a gamma globin level at least 2-fold above the negativecontrol.

[1062] The following polypeptides tested positive in this assay:PRO4352, PRO4354, PRO4408, PRO6030 and PRO4499.

Example 41 Mouse Kidney Mesangial Cell Proliferation Assay (Assay 92)

[1063] This assay shows that certain polypeptides of the invention actto induce proliferation of mammalian kidney mesangial cells and,therefore, are useful for treating kidney disorders associated withdecreased mesangial cell function such as Berger disease or othernephropathies associated with Schonlein-Henoch purpura, celiac disease,dermatitis herpetiformis or Crohn disease. The assay is performed asfollows. On day one, mouse kidney mesangial cells are plated on a 96well plate in growth media (3:1 mixture of Dulbecco's modified Eagle'smedium and Ham's F12 medium, 95% fetal bovine serum, 5% supplementedwith 14 mM HEPES) and grown overnight. On day 2, PRO polypeptides arediluted at 2 concentrations(1% and 0.1%) in serum-free medium and addedto the cells. Control samples are serum-free medium alone. On day 4, 20μl of the Cell Titer 96 Aqueous one solution reagent (Progema) was addedto each well and the colormetric reaction was allowed to proceed for 2hours. The absorbance (OD) is then measured at 490 nm. A positive in theassay is anything that gives an absorbance reading which is at least 15%above the control reading.

[1064] The following polypeptide tested positive in this assay: PRO4380,PRO4408 and PRO4425.

[1065] Deposit of Material

[1066] The following materials have been deposited with the AmericanType Culture Collection, 12301 Parklawn Drive, Rockville, Md., USA(ATCC): TABLE 7 Material ATCC Dep. No. Deposit Date DNA44686-1653 203581Jan. 12, 1999 DNA59608-2577 203870 Mar. 23, 1999 DNA62377-1381 203552Dec. 22, 1998 DNA77623-2524 203546 Dec. 22, 1998 DNA79230-2525 203549Dec. 22, 1998 DNA79862-2522 203550 Dec. 22, 1998 DNA80136-2503 203541Dec. 15, 1998 DNA80145-2594 204-PTA Jun. 8, 1999 DNA84917-2597 203863Mar. 23, 1999 DNA84920-2614 203966 Apr. 27, 1999 DNA86576-2595 203868Mar. 23, 1999 DNA87976-2593 203888 Mar. 30, 1999 DNA92234-2602 203948Apr. 20, 1999 DNA92256-2596 203891 Mar. 30, 1999 DNA92274-2617 203971Apr. 27, 1999 DNA92929-2534 203586 Jan. 12, 1999 DNA93011-2637  20-PTAMay 4, 1999 DNA96042-2682 382-PTA Jul. 20, 1999 DNA96850-2705 479-PTAAug. 3, 1999 DNA96857-2636  17-PTA May 4, 1999 DNA96867-2620 203972 Apr.27, 1999 DNA96878-2626  23-PTA May 4, 1999 DNA96899-2641 119-PTA May 25,1999

[1067] These deposits were made under the provisions of the BudapestTreaty on the International Recognition of the Deposit of Microorganismsfor the Purpose of Patent Procedure and the Regulations thereunder(Budapest Treaty). This assures maintenance of a viable culture of thedeposit for 30 years from the date of deposit. The deposits will be madeavailable by ATCC under the terms of the Budapest Treaty, and subject toan agreement between Genentech, Inc. and ATCC, which assures permanentand unrestricted availability of the progeny of the culture of thedeposit to the public upon issuance of the pertinent U.S. patent or uponlaying open to the public of any U.S. or foreign patent application,whichever comes first, and assures availability of the progeny to onedetermined by the U.S. Commissioner of Patents and Trademarks to beentitled thereto according to 35 USC § 122 and the Commissioner's rulespursuant thereto (including 37 CFR § 1.14 with particular reference to886 OG 638).

[1068] The assignee of the present application has agreed that if aculture of the materials on deposit should die or be lost or destroyedwhen cultivated under suitable conditions, the materials will bepromptly replaced on notification with another of the same. Availabilityof the deposited material is not to be construed as a license topractice the invention in contravention of the rights granted under theauthority of any government in accordance with its patent laws.

[1069] The foregoing written specification is considered to besufficient to enable one skilled in the art to practice the invention.The present invention is not to be limited in scope by the constructdeposited, since the deposited embodiment is intended as a singleillustration of certain aspects of the invention and any constructs thatare functionally equivalent are within the scope of this invention. Thedeposit of material herein does not constitute an admission that thewritten description herein contained is inadequate to enable thepractice of any aspect of the invention, including the best modethereof, nor is it to be construed as limiting the scope of the claimsto the specific illustrations that it represents. Indeed, variousmodifications of the invention in addition to those shown and describedherein will become apparent to those skilled in the art from theforegoing description and fall within the scope of the appended claims.APPENDIX A I hereby claim the benefit under Title 35, United StatesCode, §119(e) of any United States provisional applications listedbelow: 1. 60/085579 Filed May 15, 1998 2. 60/112514 Filed Dec. 15, 19983. 60/113300 Filed Dec. 22, 1998 4. 60/113430 Filed Dec. 23, 1998 5.60/113605 Filed Dec. 23, 1998 6. 60/113621 Filed Dec. 23, 1998 7.60/114140 Filed Dec. 23, 1998 8. 60/115552 Filed Jan. 12, 1999 9.60/116843 Filed Jan. 22, 1999 10. 60/125774 Filed Mar. 23, 1999 11.60/125778 Filed Mar. 23, 1999 12. 60/125826 Filed Mar. 24, 1999 13.60/127035 Filed Mar. 31, 1999 14. 60/127706 Filed Apr. 5, 1999 15.60/129122 Filed Apr. 13, 1999 16. 60/130359 Filed Apr. 21, 1999 17.60/131270 Filed Apr. 27, 1999 18. 60/131272 Filed Apr. 27, 1999 19.60/131291 Filed Apr. 27, 1999 20. 60/132371 Filed May 4, 1999 21.60/132379 Filed May 4, 1999 22. 60/132383 Filed May 4, 1999 23.60/135750 Filed May 25, 1999 24. 60/138166 Filed Jun. 8, 1999 25.60/144791 Filed Jul. 20, 1999 26. 60/146970 Filed Aug. 3, 1999 27.60/162506 Filed Oct. 29, 1999

[1070] APPENDIX B I hereby claim the benefit under Title 35, UnitedStates Code, §120 of any United States applications listed below: USFILINGS 1. 09/311832 Filed May 14, 1999 2. 09/380142 Filed Aug. 25,1999, now abandoned 3. 09/644848 Filed Aug. 22, 2000 4. 09/747259 FiledDec. 20, 2000 5. 09/816744 Filed Mar. 22, 2001 6. 09/854208 Filed May10, 2001 7. 09/854280 Filed May 10, 2001 8. 09/874503 Filed Jun. 5, 20019. 09/869599 Filed Jun. 29, 2001 10. 09/908,827 Filed Jul. 18, 2001 PCTAPPLICATIONS 1. PCT/US99/10733 Filed May 14, 1999 2. PCT/US99/28551Filed Dec. 2, 1999 3. PCT/US99/30720 Filed Dec. 22, 1999 4.PCT/US00/05601 Filed Mar. 1, 2000 5. PCT/US00/05841 Filed Mar. 2, 20006. PCT/US00/14042 Filed May 22, 2000 7. PCT/US00/15264 Filed Jun. 2,2000 8. PCT/US00/23522 Filed Aug. 23, 2000 9. PCT/US00/23328 Filed Aug.24, 2000 10. PCT/US00/32678 Filed Dec. 1, 2000 11. PCT/US00/34956 FiledDec. 20, 2000 12. PCT/US01/06520 Filed Feb. 28, 2001 13. PCT/US01/17800Filed Jun. 1, 2001 14. PCT/US01/19692 Filed Jun. 20, 2001 15.PCT/US01/21066 Filed Jun. 29, 2001 16. PCT/US01/21735 Filed Jul. 9, 2001

[1071]

1 80 1 1712 DNA Homo Sapien 1 ggcatctgcc cgaggagacc acgctcctggagctctgctg tcttctcagg 50 gagactctga ggctctgttg agaatcatgc tttggaggcagctcatctat 100 tggcaactgc tggctttgtt tttcctccct ttttgcctgt gtcaagatga150 atacatggag tctccacaaa ccggaggact acccccagac tgcagtaagt 200gttgtcatgg agactacagc tttcgaggct accaaggccc ccctgggcca 250 ccgggccctcctggcattcc aggaaaccat ggaaacaatg gcaacaatgg 300 agccactggt catgaaggagccaaaggtga gaagggcgac aaaggtgacc 350 tggggcctcg aggggagcgg gggcagcatggccccaaagg agagaagggc 400 tacccgggga ttccaccaga acttcagatt gcattcatggcttctctggc 450 aacccacttc agcaatcaga acagtgggat tatcttcagc agtgttgaga500 ccaacattgg aaacttcttt gatgtcatga ctggtagatt tggggcccca 550gtatcaggtg tgtatttctt caccttcagc atgatgaagc atgaggatgt 600 tgaggaagtgtatgtgtacc ttatgcacaa tggcaacaca gtcttcagca 650 tgtacagcta tgaaatgaagggcaaatcag atacatccag caatcatgct 700 gtgctgaagc tagccaaagg ggatgaggtttggctgcgaa tgggcaatgg 750 cgctctccat ggggaccacc aacgcttctc cacctttgcaggattcctgc 800 tctttgaaac taagtaaata tatgactaga atagctccac tttggggaag850 acttgtagct gagctgattt gttacgatct gaggaacatt aaagttgagg 900gttttacatt gctgtattca aaaaattatt ggttgcaatg ttgttcacgc 950 tacaggtacaccaataatgt tggacaattc aggggctcag aagaatcaac 1000 cacaaaatag tcttctcagatgaccttgac taatatactc agcatcttta 1050 tcactctttc cttggcacct aaaagataattctcctctga cgcaggttgg 1100 aaatattttt ttctatcaca gaagtcattt gcaaagaattttgactactc 1150 tgcttttaat ttaataccag ttttcaggaa cccctgaagt tttaagttca1200 ttattcttta taacatttga gagaatcgga tgtagtgata tgacagggct 1250ggggcaagaa caggggcact agctgcctta ttagctaatt tagtgccctc 1300 cgtgttcagcttagcctttg accctttcct tttgatccac aaaatacatt 1350 aaaactctga attcacatacaatgctattt taaagtcaat agattttagc 1400 tataaagtgc ttgaccagta atgtggttgtaattttgtgt atgttccccc 1450 acatcgcccc caacttcgga tgtggggtca ggaggttgaggttcactatt 1500 aacaaatgtc ataaatatct catagaggta cagtgccaat agatattcaa1550 atgttgcatg ttgaccagag ggattttata tctgaagaac atacactatt 1600aataaatacc ttagagaaag attttgacct ggctttagat aaaactgtgg 1650 caagaaaaatgtaatgagca atatatggaa ataaacacac ctttgttaaa 1700 gataaaaaaa aa 1712 2246 PRT Homo Sapien 2 Met Leu Trp Arg Gln Leu Ile Tyr Trp Gln Leu LeuAla Leu Phe 1 5 10 15 Phe Leu Pro Phe Cys Leu Cys Gln Asp Glu Tyr MetGlu Ser Pro 20 25 30 Gln Thr Gly Gly Leu Pro Pro Asp Cys Ser Lys Cys CysHis Gly 35 40 45 Asp Tyr Ser Phe Arg Gly Tyr Gln Gly Pro Pro Gly Pro ProGly 50 55 60 Pro Pro Gly Ile Pro Gly Asn His Gly Asn Asn Gly Asn Asn Gly65 70 75 Ala Thr Gly His Glu Gly Ala Lys Gly Glu Lys Gly Asp Lys Gly 8085 90 Asp Leu Gly Pro Arg Gly Glu Arg Gly Gln His Gly Pro Lys Gly 95 100105 Glu Lys Gly Tyr Pro Gly Ile Pro Pro Glu Leu Gln Ile Ala Phe 110 115120 Met Ala Ser Leu Ala Thr His Phe Ser Asn Gln Asn Ser Gly Ile 125 130135 Ile Phe Ser Ser Val Glu Thr Asn Ile Gly Asn Phe Phe Asp Val 140 145150 Met Thr Gly Arg Phe Gly Ala Pro Val Ser Gly Val Tyr Phe Phe 155 160165 Thr Phe Ser Met Met Lys His Glu Asp Val Glu Glu Val Tyr Val 170 175180 Tyr Leu Met His Asn Gly Asn Thr Val Phe Ser Met Tyr Ser Tyr 185 190195 Glu Met Lys Gly Lys Ser Asp Thr Ser Ser Asn His Ala Val Leu 200 205210 Lys Leu Ala Lys Gly Asp Glu Val Trp Leu Arg Met Gly Asn Gly 215 220225 Ala Leu His Gly Asp His Gln Arg Phe Ser Thr Phe Ala Gly Phe 230 235240 Leu Leu Phe Glu Thr Lys 245 3 43 DNA Artificial Sequence Syntheticoligonucleotide probe 3 tgtaaaacga cggccagtta aatagacctg caattattaa tct43 4 41 DNA Artificial Sequence Synthetic oligonucleotide probe 4caggaaacag ctatgaccac ctgcacacct gcaaatccat t 41 5 24 DNA ArtificialSequence Synthetic oligonucleotide probe 5 gcaacaatgg agccactggt catg 246 24 DNA Artificial Sequence Synthetic oligonucleotide probe 6gcaaaggtgg agaagcgttg gtgg 24 7 52 DNA Artificial Sequence Syntheticoligonucleotide probe 7 cccacttcag caatcagaac agtgggatta tctttcagcagtgtttgaga 50 cc 52 8 1579 DNA Homo Sapien 8 gagagaatag ctacagattctccatcctca gtctttgcaa ggcgacagct 50 gtgccagccg ggctctggca ggctcctggcagcatggcag tgaagcttgg 100 gaccctcctg ctggcccttg ccctgggcct ggcccagccagcctctgccc 150 gccggaagct gctggtgttt ctgctggatg gttttcgctc agactacatc200 agtgatgagg cgctggagtc attgcctggt ttcaaagaga ttgtgagcag 250gggagtaaaa gtggattact tgactccaga cttccctagt ctctcgtatc 300 ccaattattataccctaatg actggccgcc attgtgaagt ccatcagatg 350 atcgggaact acatgtgggaccccaccacc aacaagtcct ttgacattgg 400 cgtcaacaaa gacagcctaa tgcctctctggtggaatgga tcagaacctc 450 tgtgggtcac tctgaccaag gccaaaagga aggtctacatgtactactgg 500 ccaggctgtg aggttgagat tctgggtgtc agacccacct actgcctaga550 atataaaaat gtcccaacgg atatcaattt tgccaatgca gtcagcgatg 600ctcttgactc cttcaagagt ggccgggccg acctggcagc catataccat 650 gagcgcattgacgtggaagg ccaccactac gggcctgcat ctccgcagag 700 gaaagatgcc ctcaaggctgtagacactgt cctgaagtac atgaccaagt 750 ggatccagga gcggggcctg caggaccgcctgaacgtcat tattttctcg 800 gatcacggaa tgaccgacat tttctggatg gacaaagtgattgagctgaa 850 taagtacatc agcctgaatg acctgcagca agtgaaggac cgcgggcctg900 ttgtgagcct ttggccggcc cctgggaaac actctgagat atataacaaa 950ctgagcacag tggaacacat gactgtctac gagaaagaag ccatcccaag 1000 caggttctattacaagaaag gaaagtttgt ctctcctttg actttagtgg 1050 ctgatgaagg ctggttcataactgagaatc gagagatgct tccgttttgg 1100 atgaacagca ccggcaggcg ggaaggttggcagcgtggat ggcacggcta 1150 cgacaacgag ctcatggaca tgcggggcat cttcctggccttcggacctg 1200 atttcaaatc caacttcaga gctgctccta tcaggtcggt ggacgtctac1250 aatgtcatgt gcaatgtggt gggcatcacc ccgctgccca acaacggatc 1300ctggtccagg gtgatgtgca tgctgaaggg ccgcgccggc actgccccgc 1350 ctgtctggcccagccactgt gccctggcac tgattcttct cttcctgctt 1400 gcataactga tcatattgcttgtctcagaa aaaaacacca tcagcaaagt 1450 gggcctccaa agccagatga ttttcattttatgtgtgaat aatagcttca 1500 ttaacacaat caagaccatg cacattgtaa atacattattcttggataat 1550 tctatacata aaagttccta cttgttaaa 1579 9 440 PRT HomoSapien 9 Met Ala Val Lys Leu Gly Thr Leu Leu Leu Ala Leu Ala Leu Gly 1 510 15 Leu Ala Gln Pro Ala Ser Ala Arg Arg Lys Leu Leu Val Phe Leu 20 2530 Leu Asp Gly Phe Arg Ser Asp Tyr Ile Ser Asp Glu Ala Leu Glu 35 40 45Ser Leu Pro Gly Phe Lys Glu Ile Val Ser Arg Gly Val Lys Val 50 55 60 AspTyr Leu Thr Pro Asp Phe Pro Ser Leu Ser Tyr Pro Asn Tyr 65 70 75 Tyr ThrLeu Met Thr Gly Arg His Cys Glu Val His Gln Met Ile 80 85 90 Gly Asn TyrMet Trp Asp Pro Thr Thr Asn Lys Ser Phe Asp Ile 95 100 105 Gly Val AsnLys Asp Ser Leu Met Pro Leu Trp Trp Asn Gly Ser 110 115 120 Glu Pro LeuTrp Val Thr Leu Thr Lys Ala Lys Arg Lys Val Tyr 125 130 135 Met Tyr TyrTrp Pro Gly Cys Glu Val Glu Ile Leu Gly Val Arg 140 145 150 Pro Thr TyrCys Leu Glu Tyr Lys Asn Val Pro Thr Asp Ile Asn 155 160 165 Phe Ala AsnAla Val Ser Asp Ala Leu Asp Ser Phe Lys Ser Gly 170 175 180 Arg Ala AspLeu Ala Ala Ile Tyr His Glu Arg Ile Asp Val Glu 185 190 195 Gly His HisTyr Gly Pro Ala Ser Pro Gln Arg Lys Asp Ala Leu 200 205 210 Lys Ala ValAsp Thr Val Leu Lys Tyr Met Thr Lys Trp Ile Gln 215 220 225 Glu Arg GlyLeu Gln Asp Arg Leu Asn Val Ile Ile Phe Ser Asp 230 235 240 His Gly MetThr Asp Ile Phe Trp Met Asp Lys Val Ile Glu Leu 245 250 255 Asn Lys TyrIle Ser Leu Asn Asp Leu Gln Gln Val Lys Asp Arg 260 265 270 Gly Pro ValVal Ser Leu Trp Pro Ala Pro Gly Lys His Ser Glu 275 280 285 Ile Tyr AsnLys Leu Ser Thr Val Glu His Met Thr Val Tyr Glu 290 295 300 Lys Glu AlaIle Pro Ser Arg Phe Tyr Tyr Lys Lys Gly Lys Phe 305 310 315 Val Ser ProLeu Thr Leu Val Ala Asp Glu Gly Trp Phe Ile Thr 320 325 330 Glu Asn ArgGlu Met Leu Pro Phe Trp Met Asn Ser Thr Gly Arg 335 340 345 Arg Glu GlyTrp Gln Arg Gly Trp His Gly Tyr Asp Asn Glu Leu 350 355 360 Met Asp MetArg Gly Ile Phe Leu Ala Phe Gly Pro Asp Phe Lys 365 370 375 Ser Asn PheArg Ala Ala Pro Ile Arg Ser Val Asp Val Tyr Asn 380 385 390 Val Met CysAsn Val Val Gly Ile Thr Pro Leu Pro Asn Asn Gly 395 400 405 Ser Trp SerArg Val Met Cys Met Leu Lys Gly Arg Ala Gly Thr 410 415 420 Ala Pro ProVal Trp Pro Ser His Cys Ala Leu Ala Leu Ile Leu 425 430 435 Leu Phe LeuLeu Ala 440 10 1047 DNA Homo Sapien 10 gccaggtgtg caggccgctc caagcccagcctgccccgct gccgccacca 50 tgacgctcct ccccggcctc ctgtttctga cctggctgcacacatgcctg 100 gcccaccatg acccctccct cagggggcac ccccacagtc acggtacccc150 acactgctac tcggctgagg aactgcccct cggccaggcc cccccacacc 200tgctggctcg aggtgccaag tgggggcagg ctttgcctgt agccctggtg 250 tccagcctggaggcagcaag ccacaggggg aggcacgaga ggccctcagc 300 tacgacccag tgcccggtgctgcggccgga ggaggtgttg gaggcagaca 350 cccaccagcg ctccatctca ccctggagataccgtgtgga cacggatgag 400 gaccgctatc cacagaagct ggccttcgcc gagtgcctgtgcagaggctg 450 tatcgatgca cggacgggcc gcgagacagc tgcgctcaac tccgtgcggc500 tgctccagag cctgctggtg ctgcgccgcc ggccctgctc ccgcgacggc 550tcggggctcc ccacacctgg ggcctttgcc ttccacaccg agttcatcca 600 cgtccccgtcggctgcacct gcgtgctgcc ccgttcagtg tgaccgccga 650 ggccgtgggg cccctagactggacacgtgt gctccccaga gggcaccccc 700 tatttatgtg tatttattgt tatttatatgcctcccccaa cactaccctt 750 ggggtctggg cattccccgt gtctggagga cagccccccactgttctcct 800 catctccagc ctcagtagtt gggggtagaa ggagctcagc acctcttcca850 gcccttaaag ctgcagaaaa ggtgtcacac ggctgcctgt accttggctc 900cctgtcctgc tcccggcttc ccttacccta tcactggcct caggccccgc 950 aggctgcctcttcccaacct ccttggaagt acccctgttt cttaaacaat 1000 tatttaagtg tacgtgtattattaaactga tgaacacatc cccaaaa 1047 11 197 PRT Homo Sapien 11 Met Thr LeuLeu Pro Gly Leu Leu Phe Leu Thr Trp Leu His Thr 1 5 10 15 Cys Leu AlaHis His Asp Pro Ser Leu Arg Gly His Pro His Ser 20 25 30 His Gly Thr ProHis Cys Tyr Ser Ala Glu Glu Leu Pro Leu Gly 35 40 45 Gln Ala Pro Pro HisLeu Leu Ala Arg Gly Ala Lys Trp Gly Gln 50 55 60 Ala Leu Pro Val Ala LeuVal Ser Ser Leu Glu Ala Ala Ser His 65 70 75 Arg Gly Arg His Glu Arg ProSer Ala Thr Thr Gln Cys Pro Val 80 85 90 Leu Arg Pro Glu Glu Val Leu GluAla Asp Thr His Gln Arg Ser 95 100 105 Ile Ser Pro Trp Arg Tyr Arg ValAsp Thr Asp Glu Asp Arg Tyr 110 115 120 Pro Gln Lys Leu Ala Phe Ala GluCys Leu Cys Arg Gly Cys Ile 125 130 135 Asp Ala Arg Thr Gly Arg Glu ThrAla Ala Leu Asn Ser Val Arg 140 145 150 Leu Leu Gln Ser Leu Leu Val LeuArg Arg Arg Pro Cys Ser Arg 155 160 165 Asp Gly Ser Gly Leu Pro Thr ProGly Ala Phe Ala Phe His Thr 170 175 180 Glu Phe Ile His Val Pro Val GlyCys Thr Cys Val Leu Pro Arg 185 190 195 Ser Val 12 24 DNA ArtificialSequence Synthetic oligonucleotide probe 12 atccacagaa gctggccttc gccg24 13 24 DNA Artificial Sequence Synthetic oligonucleotide probe 13gggacgtgga tgaactcggt gtgg 24 14 40 DNA Artificial Sequence Syntheticoligonucleotide probe 14 tatccacaga agctggcctt cgccgagtgc ctgtgcagag 4015 660 DNA Homo Sapien 15 cggccagggc gccgacagcc cgacctcacc aggagaacatgcagctcggc 50 actgggctcc tgctggccgc cgtcctgagc ctgcagctgg ctgcagccga 100agccatatgg tgtcaccagt gcacgggctt cggagggtgc tcccatggat 150 ccagatgcctgagggactcc acccactgtg tcaccactgc cacccgggtc 200 ctcagcaaca ccgaggatttgcctctggtc accaagatgt gccacatagg 250 ctgccccgat atccccagcc tgggcctgggcccctacgta tccatcgctt 300 gctgccagac cagcctctgc aaccatgact gacggctgccctcctccagg 350 cccccggacg ctcagccccc acagccccca cagcctggcg ccagggctca400 cggccgcccc tccctcgaga ctggccagcc cacctctccc ggcctctgca 450gccaccgtcc agcaccgctt gtcctaggga agtcctgcgt ggagtcttgc 500 ctcaatctgctgccgtccaa gcctggggcc catcgtgcct gccgcccctt 550 caggtcccga cctccccacaataaaatgtg attggatcgt gtggtacaaa 600 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa 650 aaaaaaaaaa 660 16 97 PRT Homo Sapien 16 MetGln Leu Gly Thr Gly Leu Leu Leu Ala Ala Val Leu Ser Leu 1 5 10 15 GlnLeu Ala Ala Ala Glu Ala Ile Trp Cys His Gln Cys Thr Gly 20 25 30 Phe GlyGly Cys Ser His Gly Ser Arg Cys Leu Arg Asp Ser Thr 35 40 45 His Cys ValThr Thr Ala Thr Arg Val Leu Ser Asn Thr Glu Asp 50 55 60 Leu Pro Leu ValThr Lys Met Cys His Ile Gly Cys Pro Asp Ile 65 70 75 Pro Ser Leu Gly LeuGly Pro Tyr Val Ser Ile Ala Cys Cys Gln 80 85 90 Thr Ser Leu Cys Asn HisAsp 95 17 2570 DNA Homo Sapien 17 ccaggaccag ggcgcaccgg ctcagcctctcacttgtcag aggccgggga 50 agagaagcaa agcgcaacgg tgtggtccaa gccggggcttctgcttcgcc 100 tctaggacat acacgggacc ccctaacttc agtcccccaa acgcgcaccc150 tcgaagtctt gaactccagc cccgcacatc cacgcgcggc acaggcgcgg 200caggcggcag gtcccggccg aaggcgatgc gcgcaggggg tcgggcagct 250 gggctcgggcggcgggagta gggcccggca gggaggcagg gaggctgcat 300 attcagagtc gcgggctgcgccctgggcag aggccgccct cgctccacgc 350 aacacctgct gctgccaccg cgccgcgatgagccgcgtgg tctcgctgct 400 gctgggcgcc gcgctgctct gcggccacgg agccttctgccgccgcgtgg 450 tcagcggcca aaaggtgtgt tttgctgact tcaagcatcc ctgctacaaa500 atggcctact tccatgaact gtccagccga gtgagctttc aggaggcacg 550cctggcttgt gagagtgagg gaggagtcct cctcagcctt gagaatgaag 600 cagaacagaagttaatagag agcatgttgc aaaacctgac aaaacccggg 650 acagggattt ctgatggtgatttctggata gggctttgga ggaatggaga 700 tgggcaaaca tctggtgcct gcccagatctctaccagtgg tctgatggaa 750 gcaattccca gtaccgaaac tggtacacag atgaaccttcctgcggaagt 800 gaaaagtgtg ttgtgatgta tcaccaacca actgccaatc ctggccttgg850 gggtccctac ctttaccagt ggaatgatga caggtgtaac atgaagcaca 900attatatttg caagtatgaa ccagagatta atccaacagc ccctgtagaa 950 aagccttatcttacaaatca accaggagac acccatcaga atgtggttgt 1000 tactgaagca ggtataattcccaatctaat ttatgttgtt ataccaacaa 1050 tacccctgct cttactgata ctggttgcttttggaacctg ttgtttccag 1100 atgctgcata aaagtaaagg aagaacaaaa actagtccaaaccagtctac 1150 actgtggatt tcaaagagta ccagaaaaga aagtggcatg gaagtataat1200 aactcattga cttggttcca gaattttgta attctggatc tgtataagga 1250atggcatcag aacaatagct tggaatggct tgaaatcaca aaggatctgc 1300 aagatgaactgtaagctccc ccttgaggca aatattaaag taatttttat 1350 atgtctatta tttcatttaaagaatatgct gtgctaataa tggagtgaga 1400 catgcttatt ttgctaaagg atgcacccaaacttcaaact tcaagcaaat 1450 gaaatggaca atgcagataa agttgttatc aacacgtcgggagtatgtgt 1500 gttagaagca attcctttta tttctttcac ctttcataag ttgttatcta1550 gtcaatgtaa tgtatattgt attgaaattt acagtgtgca aaagtatttt 1600acctttgcat aagtgtttga taaaaatgaa ctgttctaat atttattttt 1650 atggcatctcatttttcaat acatgctctt ttgattaaag aaacttatta 1700 ctgttgtcaa ctgaattcacacacacacaa atatagtacc atagaaaaag 1750 tttgttttct cgaaataatt catctttcagcttctctgct tttggtcaat 1800 gtctaggaaa tctcttcaga aataagaagc tatttcattaagtgtgatat 1850 aaacctcctc aaacatttta cttagaggca aggattgtct aatttcaatt1900 gtgcaagaca tgtgccttat aattattttt agcttaaaat taaacagatt 1950ttgtaataat gtaactttgt taataggtgc ataaacacta atgcagtcaa 2000 tttgaacaaaagaagtgaca tacacaatat aaatcatatg tcttcacacg 2050 ttgcctatat aatgagaagcagctctctga gggttctgaa atcaatgtgg 2100 tccctctctt gcccactaaa caaagatggttgttcggggt ttgggattga 2150 cactggaggc agatagttgc aaagttagtc taaggtttccctagctgtat 2200 ttagcctctg actatattag tatacaaaga ggtcatgtgg ttgagaccag2250 gtgaatagtc actatcagtg tggagacaag cacagcacac agacatttta 2300ggaaggaaag gaactacgaa atcgtgtgaa aatgggttgg aacccatcag 2350 tgatcgcatattcattgatg agggtttgct tgagatagaa aatggtggct 2400 cctttctgtc ttatctcctagtttcttcaa tgcttacgcc ttgttcttct 2450 caagagaaag ttgtaactct ctggtcttcatatgtccctg tgctcctttt 2500 aaccaaataa agagttcttg tttctggggg aaaaaaaaaaaaaaaaaaaa 2550 aaaaaaaaaa aaaaaaaaaa 2570 18 273 PRT Homo Sapien 18 MetSer Arg Val Val Ser Leu Leu Leu Gly Ala Ala Leu Leu Cys 1 5 10 15 GlyHis Gly Ala Phe Cys Arg Arg Val Val Ser Gly Gln Lys Val 20 25 30 Cys PheAla Asp Phe Lys His Pro Cys Tyr Lys Met Ala Tyr Phe 35 40 45 His Glu LeuSer Ser Arg Val Ser Phe Gln Glu Ala Arg Leu Ala 50 55 60 Cys Glu Ser GluGly Gly Val Leu Leu Ser Leu Glu Asn Glu Ala 65 70 75 Glu Gln Lys Leu IleGlu Ser Met Leu Gln Asn Leu Thr Lys Pro 80 85 90 Gly Thr Gly Ile Ser AspGly Asp Phe Trp Ile Gly Leu Trp Arg 95 100 105 Asn Gly Asp Gly Gln ThrSer Gly Ala Cys Pro Asp Leu Tyr Gln 110 115 120 Trp Ser Asp Gly Ser AsnSer Gln Tyr Arg Asn Trp Tyr Thr Asp 125 130 135 Glu Pro Ser Cys Gly SerGlu Lys Cys Val Val Met Tyr His Gln 140 145 150 Pro Thr Ala Asn Pro GlyLeu Gly Gly Pro Tyr Leu Tyr Gln Trp 155 160 165 Asn Asp Asp Arg Cys AsnMet Lys His Asn Tyr Ile Cys Lys Tyr 170 175 180 Glu Pro Glu Ile Asn ProThr Ala Pro Val Glu Lys Pro Tyr Leu 185 190 195 Thr Asn Gln Pro Gly AspThr His Gln Asn Val Val Val Thr Glu 200 205 210 Ala Gly Ile Ile Pro AsnLeu Ile Tyr Val Val Ile Pro Thr Ile 215 220 225 Pro Leu Leu Leu Leu IleLeu Val Ala Phe Gly Thr Cys Cys Phe 230 235 240 Gln Met Leu His Lys SerLys Gly Arg Thr Lys Thr Ser Pro Asn 245 250 255 Gln Ser Thr Leu Trp IleSer Lys Ser Thr Arg Lys Glu Ser Gly 260 265 270 Met Glu Val 19 24 DNAArtificial Sequence Synthetic oligonucleotide probe 19 caccaaccaactgccaatcc tggc 24 20 26 DNA Artificial Sequence Syntheticoligonucleotide probe 20 accacattct gatgggtgtc tcctgg 26 21 49 DNAArtificial Sequence Synthetic oligonucleotide probe 21 gggtccctacctttaccagt ggaatgatga caggtgtaac atgaagcac 49 22 3824 DNA Homo Sapien 22ggagaatgga gagagcagtg agagtggagt ccggggtcct ggtcggggtg 50 gtctgtctgctcctggcatg ccctgccaca gccactgggc ccgaagttgc 100 tcagcctgaa gtagacaccaccctgggtcg tgtgcgaggc cggcaggtgg 150 gcgtgaaggg cacagaccgc cttgtgaatgtctttctggg cattccattt 200 gcccagccgc cactgggccc tgaccggttc tcagccccacacccagcaca 250 gccctgggag ggtgtgcggg atgccagcac tgcgccccca atgtgcctac300 aagacgtgga gagcatgaac agcagcagat ttgtcctcaa cggaaaacag 350cagatcttct ccgtttcaga ggactgcctg gtcctcaacg tctatagccc 400 agctgaggtccccgcagggt ccggtaggcc ggtcatggta tgggtccatg 450 gaggcgctct gataactggcgctgccacct cctacgatgg atcagctctg 500 gctgcctatg gggatgtggt cgtggttacagtccagtacc gccttggggt 550 ccttggcttc ttcagcactg gagatgagca tgcacctggcaaccagggct 600 tcctagatgt ggtagctgct ttgcgctggg tgcaagaaaa catcgccccc650 ttcgggggtg acctcaactg tgtcactgtc tttggtggat ctgccggtgg 700gagcatcatc tctggcctgg tcctgtcccc agtggctgca gggctgttcc 750 acagagccatcacacagagt ggggtcatca ccaccccagg gatcatcgac 800 tctcaccctt ggcccctagctcagaaaatc gcaaacacct tggcctgcag 850 ctccagctcc ccggctgaga tggtgcagtgccttcagcag aaagaaggag 900 aagagctggt ccttagcaag aagctgaaaa atactatctatcctctcacc 950 gttgatggca ctgtcttccc caaaagcccc aaggaactcc tgaaggagaa1000 gcccttccac tctgtgccct tcctcatggg tgtcaacaac catgagttca 1050gctggctcat ccccaggggc tggggtctcc tggatacaat ggagcagatg 1100 agccgggaggacatgctggc catctcaaca cccgtcttga ccagtctgga 1150 tgtgccccct gagatgatgcccaccgtcat agatgaatac ctaggaagca 1200 actcggacgc acaagccaaa tgccaggcgttccaggaatt catgggtgac 1250 gtattcatca atgttcccac cgtcagtttt tcaagataccttcgagattc 1300 tggaagccct gtctttttct atgagttcca gcatcgaccc agttcttttg1350 cgaagatcaa acctgcctgg gtgaaggctg atcatggggc cgagggtgct 1400tttgtgttcg gaggtccctt cctcatggac gagagctccc gcctggcctt 1450 tccagaggccacagaggagg agaagcagct aagcctcacc atgatggccc 1500 agtggaccca ctttgcccggacaggggacc ccaatagcaa ggctctgcct 1550 ccttggcccc aattcaacca ggcggaacaatatctggaga tcaacccagt 1600 gccacgggcc ggacagaagt tcagggaggc ctggatgcagttctggtcag 1650 agacgctccc cagcaagata caacagtggc accagaagca gaagaacagg1700 aaggcccagg aggacctctg aggccaggcc tgaaccttct tggctggggc 1750aaaccactct tcaagtggtg gcagagtccc agcacggcag cccgcctctc 1800 cccctgctgagactttaatc tccaccagcc cttaaagtgt cggccgctct 1850 gtgactggag ttatgctcttttgaaatgtc acaaggccgc ctcccacctc 1900 tggggcattg tacaagttct tccctctccctgaagtgcct ttcctgcttt 1950 cttcgtggta ggttctagca cattcctcta gcttcctggaggactcactc 2000 cccaggaagc cttccctgcc ttctctgggc tgtgcggccc cgagtctgcg2050 tccattagag cacagtccac ccgaggctag caccgtgtct gtgtctgtct 2100ccccctcaga ggagctctct caaaatgggg attagcctaa ccccactctg 2150 tcacccacaccaggatcggg tgggacctgg agctaggggg tgtttgctga 2200 gtgagtgagt gaaacacagaatatgggaat ggcagctgct gaacttgaac 2250 ccagagcctt caggtgccaa agccatactcaggcccccac cgacattgtc 2300 caccctggcc agaagggtgc atgccaatgg cagagacctgggatgggaga 2350 agtcctgggg cgccagggga tccagcctag agcagacctt agcccctgac2400 taaggcctca gactagggcg ggaggggtct cctcctctct gctgcccagt 2450cctggcccct gcacaagaca acagaatcca tcagggccat gagtgtcacc 2500 cagacctgaccctcaccaat tccagcccct gaccctcagg acgctggatg 2550 ccagctccca gccccagtgccgggtcctcc ctcccttcct ggcttgggga 2600 gaccagtttc tggggagctt ccaagagcacccaccaagac acagcaggac 2650 aggccagggg agggcatctg gaccagggca tccgtcgggctattgtcaca 2700 gagaaaagaa gagacccacc cactcgggct gcaaaaggtg aaaagcacca2750 agaggttttc agatggaagt gagaggtgac agtgtgctgg cagccctcac 2800agccctcgct tgctctccct gccgcctctg cctgggctcc cactttggca 2850 gcacttgaggagcccttcaa cccgccgctg cactgtagga gcccctttct 2900 gggctggcca aggccggagccagctccctc agcttgcggg gaggtgcgga 2950 gggagagggg cgggcaggaa ccggggctgcgcgcagcgct tgcgggccag 3000 agtgagttcc gggtgggcgt gggctcggcg gggccccactcagagcagct 3050 ggccggcccc aggcagtgag ggccttagca cctgggccag cagctgctgt3100 gctcgatttc tcgctgggcc ttagctgcct ccccgcgggg cagggctcgg 3150gacctgcagc cctccatgcc tgaccctccc cccacccccc gtgggctcct 3200 gtgcggccggagcctcccca aggagcgccg ccccctgctc cacagcgccc 3250 agtcccatcg accacccaagggctgaggag tgcgggtgca cagcgcggga 3300 ctggcaggca gctccacctg ctgccccagtgctggatcca ctgggtgaag 3350 ccagctgggc tcctgagtct ggtggggact tggagaacctttatgtctag 3400 ctaagggatt gtaaatacac cgatgggcac tctgtatcta gctcaaggtt3450 tgtaaacaca ccaatcagca ccctgtgtct agctcagtgt ttgtgaatgc 3500accaatccac actctgtatc tggctactct ggtggggact tggagaacct 3550 ttgtgtccacactctgtatc tagctaatct agtggggatg tggagaacct 3600 ttgtgtctag ctcagggatcgtaaacgcac caatcagcac cctgtcaaaa 3650 cagaccactt gactctctgt aaaatggaccaatcagcagg atgtgggtgg 3700 ggcgagacaa gagaataaaa gcaggctgcc tgagccagcagtgacaaccc 3750 ccctcgggtc ccctcccacg ccgtggaagc tttgttcttt cgctctttgc3800 aataaatctt gctactgccc aaaa 3824 23 571 PRT Homo Sapien 23 Met GluArg Ala Val Arg Val Glu Ser Gly Val Leu Val Gly Val 1 5 10 15 Val CysLeu Leu Leu Ala Cys Pro Ala Thr Ala Thr Gly Pro Glu 20 25 30 Val Ala GlnPro Glu Val Asp Thr Thr Leu Gly Arg Val Arg Gly 35 40 45 Arg Gln Val GlyVal Lys Gly Thr Asp Arg Leu Val Asn Val Phe 50 55 60 Leu Gly Ile Pro PheAla Gln Pro Pro Leu Gly Pro Asp Arg Phe 65 70 75 Ser Ala Pro His Pro AlaGln Pro Trp Glu Gly Val Arg Asp Ala 80 85 90 Ser Thr Ala Pro Pro Met CysLeu Gln Asp Val Glu Ser Met Asn 95 100 105 Ser Ser Arg Phe Val Leu AsnGly Lys Gln Gln Ile Phe Ser Val 110 115 120 Ser Glu Asp Cys Leu Val LeuAsn Val Tyr Ser Pro Ala Glu Val 125 130 135 Pro Ala Gly Ser Gly Arg ProVal Met Val Trp Val His Gly Gly 140 145 150 Ala Leu Ile Thr Gly Ala AlaThr Ser Tyr Asp Gly Ser Ala Leu 155 160 165 Ala Ala Tyr Gly Asp Val ValVal Val Thr Val Gln Tyr Arg Leu 170 175 180 Gly Val Leu Gly Phe Phe SerThr Gly Asp Glu His Ala Pro Gly 185 190 195 Asn Gln Gly Phe Leu Asp ValVal Ala Ala Leu Arg Trp Val Gln 200 205 210 Glu Asn Ile Ala Pro Phe GlyGly Asp Leu Asn Cys Val Thr Val 215 220 225 Phe Gly Gly Ser Ala Gly GlySer Ile Ile Ser Gly Leu Val Leu 230 235 240 Ser Pro Val Ala Ala Gly LeuPhe His Arg Ala Ile Thr Gln Ser 245 250 255 Gly Val Ile Thr Thr Pro GlyIle Ile Asp Ser His Pro Trp Pro 260 265 270 Leu Ala Gln Lys Ile Ala AsnThr Leu Ala Cys Ser Ser Ser Ser 275 280 285 Pro Ala Glu Met Val Gln CysLeu Gln Gln Lys Glu Gly Glu Glu 290 295 300 Leu Val Leu Ser Lys Lys LeuLys Asn Thr Ile Tyr Pro Leu Thr 305 310 315 Val Asp Gly Thr Val Phe ProLys Ser Pro Lys Glu Leu Leu Lys 320 325 330 Glu Lys Pro Phe His Ser ValPro Phe Leu Met Gly Val Asn Asn 335 340 345 His Glu Phe Ser Trp Leu IlePro Arg Gly Trp Gly Leu Leu Asp 350 355 360 Thr Met Glu Gln Met Ser ArgGlu Asp Met Leu Ala Ile Ser Thr 365 370 375 Pro Val Leu Thr Ser Leu AspVal Pro Pro Glu Met Met Pro Thr 380 385 390 Val Ile Asp Glu Tyr Leu GlySer Asn Ser Asp Ala Gln Ala Lys 395 400 405 Cys Gln Ala Phe Gln Glu PheMet Gly Asp Val Phe Ile Asn Val 410 415 420 Pro Thr Val Ser Phe Ser ArgTyr Leu Arg Asp Ser Gly Ser Pro 425 430 435 Val Phe Phe Tyr Glu Phe GlnHis Arg Pro Ser Ser Phe Ala Lys 440 445 450 Ile Lys Pro Ala Trp Val LysAla Asp His Gly Ala Glu Gly Ala 455 460 465 Phe Val Phe Gly Gly Pro PheLeu Met Asp Glu Ser Ser Arg Leu 470 475 480 Ala Phe Pro Glu Ala Thr GluGlu Glu Lys Gln Leu Ser Leu Thr 485 490 495 Met Met Ala Gln Trp Thr HisPhe Ala Arg Thr Gly Asp Pro Asn 500 505 510 Ser Lys Ala Leu Pro Pro TrpPro Gln Phe Asn Gln Ala Glu Gln 515 520 525 Tyr Leu Glu Ile Asn Pro ValPro Arg Ala Gly Gln Lys Phe Arg 530 535 540 Glu Ala Trp Met Gln Phe TrpSer Glu Thr Leu Pro Ser Lys Ile 545 550 555 Gln Gln Trp His Gln Lys GlnLys Asn Arg Lys Ala Gln Glu Asp 560 565 570 Leu 24 22 DNA ArtificialSequence Synthetic oligonucleotide probe 24 gcaaagctct gcctccttgg cc 2225 25 DNA Artificial Sequence Synthetic oligonucleotide probe 25gggtggactg tgctctaatg gacgc 25 26 18 DNA Artificial Sequence Syntheticoligonucleotide probe 26 cgtggcactg ggttgatc 18 27 45 DNA ArtificialSequence Synthetic oligonucleotide probe 27 gatgcagttc tggtcagagacgctccccag caagatacaa cagtg 45 28 1342 DNA Homo Sapien 28 catggagcctcttgcagctt acccgctaaa atgttccggg cccagagcaa 50 aggtatttgc agttttgctgtctatagttc tatgcacagt aacgctattt 100 cttctacaac taaaattcct caaacctaaaatcaacagct tttatgcctt 150 tgaagtgaag gatgcaaaag gaagaactgt ttctctggaaaagtataaag 200 gcaaagtttc actagttgta aacgtggcca gtgactgcca actcacagac250 agaaattact tagggctgaa ggaactgcac aaagagtttg gaccatccca 300cttcagcgtg ttggcttttc cctgcaatca gtttggagaa tcggagcccc 350 gcccaagcaaggaagtagaa tcttttgcaa gaaaaaacta cggagtaact 400 ttccccatct tccacaagattaagattcta ggatctgaag gagaacctgc 450 atttagattt cttgttgatt cttcaaagaaggaaccaagg tggaattttt 500 ggaagtatct tgtcaaccct gagggtcaag ttgtgaagttctggaggcca 550 gaggagccca ttgaagtcat caggcctgac atagcagctc tggttagaca600 agtgatcata aaaaagaaag aggatctatg agaatgccat tgcgtttcta 650atagaacaga gaaatgtctc catgagggtt tggtctcatt ttaaacattt 700 tttttttggagacagtgtct cactctgtca cccaggctgg agtgcagtag 750 tgcgttctca gctcattgcaacctctgcct ttttaaacat gctattaaat 800 gtggcaatga aggatttttt tttaatgttatcttgctatt aagtggtaat 850 gaatgttccc aggatgagga tgttacccaa agcaaaaatcaagagtagcc 900 aaagaatcaa catgaaatat attaactact tcctctgacc atactaaaga950 attcagaata cacagtgacc aatgtgcctc aatatcttat tgttcaactt 1000gacattttct aggactgtac ttgatgaaaa tgccaacaca ctagaccact 1050 ctttggattcaagagcactg tgtatgactg aaatttctgg aataactgta 1100 aatggttatg ttaatggaataaaacacaaa tgttgaaaaa tgtaaaatat 1150 atatacatag attcaaatcc ttatatatgtatgcttgttt tgtgtacagg 1200 attttgtttt ttctttttaa gtacaggttc ctagtgttttactataactg 1250 tcactatgta tgtaactgac atatataaat agtcatttat aaatgaccgt1300 attataacat ttgaaaaagt cttcatcaaa aaaaaaaaaa aa 1342 29 209 PRT HomoSapien 29 Met Glu Pro Leu Ala Ala Tyr Pro Leu Lys Cys Ser Gly Pro Arg 15 10 15 Ala Lys Val Phe Ala Val Leu Leu Ser Ile Val Leu Cys Thr Val 2025 30 Thr Leu Phe Leu Leu Gln Leu Lys Phe Leu Lys Pro Lys Ile Asn 35 4045 Ser Phe Tyr Ala Phe Glu Val Lys Asp Ala Lys Gly Arg Thr Val 50 55 60Ser Leu Glu Lys Tyr Lys Gly Lys Val Ser Leu Val Val Asn Val 65 70 75 AlaSer Asp Cys Gln Leu Thr Asp Arg Asn Tyr Leu Gly Leu Lys 80 85 90 Glu LeuHis Lys Glu Phe Gly Pro Ser His Phe Ser Val Leu Ala 95 100 105 Phe ProCys Asn Gln Phe Gly Glu Ser Glu Pro Arg Pro Ser Lys 110 115 120 Glu ValGlu Ser Phe Ala Arg Lys Asn Tyr Gly Val Thr Phe Pro 125 130 135 Ile PheHis Lys Ile Lys Ile Leu Gly Ser Glu Gly Glu Pro Ala 140 145 150 Phe ArgPhe Leu Val Asp Ser Ser Lys Lys Glu Pro Arg Trp Asn 155 160 165 Phe TrpLys Tyr Leu Val Asn Pro Glu Gly Gln Val Val Lys Phe 170 175 180 Trp ArgPro Glu Glu Pro Ile Glu Val Ile Arg Pro Asp Ile Ala 185 190 195 Ala LeuVal Arg Gln Val Ile Ile Lys Lys Lys Glu Asp Leu 200 205 30 24 DNAArtificial Sequence Synthetic oligonucleotide probe 30 atcctccaacatggagcctc ttgc 24 31 20 DNA Artificial Sequence Syntheticoligonucleotide probe 31 gtatcttgtc aaccctgagg 20 32 24 DNA ArtificialSequence Synthetic oligonucleotide probe 32 taaccagagc tgctatgtca ggcc24 33 50 DNA Artificial Sequence Synthetic oligonucleotide probe 33aggcaaagtt tcactagttg taaacgtggc cagtgactgc caactcacag 50 34 3721 DNAHomo Sapien 34 tgtcgcctgg ccctcgccat gcagaccccg cgagcgtccc ctccccgccc 50ggccctcctg cttctgctgc tgctactggg gggcgcccac ggcctctttc 100 ctgaggagccgccgccgctt agcgtggccc ccagggacta cctgaaccac 150 tatcccgtgt ttgtgggcagcgggcccgga cgcctgaccc ccgcagaagg 200 tgctgacgac ctcaacatcc agcgagtcctgcgggtcaac aggacgctgt 250 tcattgggga cagggacaac ctctaccgcg tagagctggagccccccacg 300 tccacggagc tgcggtacca gaggaagctg acctggagat ctaaccccag350 cgacataaac gtgtgtcgga tgaagggcaa acaggagggc gagtgtcgaa 400acttcgtaaa ggtgctgctc cttcgggacg agtccacgct ctttgtgtgc 450 ggttccaacgccttcaaccc ggtgtgcgcc aactacagca tagacaccct 500 gcagcccgtc ggagacaacatcagcggtat ggcccgctgc ccgtacgacc 550 ccaagcacgc caatgttgcc ctcttctctgacgggatgct cttcacagct 600 actgttaccg acttcctagc cattgatgct gtcatctaccgcagcctcgg 650 ggacaggccc accctgcgca ccgtgaaaca tgactccaag tggttcaaag700 agccttactt tgtccatgcg gtggagtggg gcagccatgt ctacttcttc 750ttccgggaga ttgcgatgga gtttaactac ctggagaagg tggtggtgtc 800 ccgcgtggcccgagtgtgca agaacgacgt gggaggctcc ccccgcgtgc 850 tggagaagca gtggacgtccttcctgaagg cgcggctcaa ctgctctgta 900 cccggagact cccatttcta cttcaacgtgctgcaggctg tcacgggcgt 950 ggtcagcctc gggggccggc ccgtggtcct ggccgttttttccacgccca 1000 gcaacagcat ccctggctcg gctgtctgcg cctttgacct gacacaggtg1050 gcagctgtgt ttgaaggccg cttccgagag cagaagtccc ccgagtccat 1100ctggacgccg gtgccggagg atcaggtgcc tcgaccccgg cccgggtgct 1150 gcgcagcccccgggatgcag tacaatgcct ccagcgcctt gccggatgac 1200 atcctcaact ttgtcaagacccaccctctg atggacgagg cggtgccctc 1250 gctgggccat gcgccctgga tcctgcggaccctgatgagg caccagctga 1300 ctcgagtggc tgtggacgtg ggagccggcc cctggggcaaccagaccgtt 1350 gtcttcctgg gttctgaggc ggggacggtc ctcaagttcc tcgtccggcc1400 caatgccagc acctcaggga cgtctgggct cagtgtcttc ctggaggagt 1450ttgagaccta ccggccggac aggtgtggac ggcccggcgg tggcgagaca 1500 gggcagcggctgctgagctt ggagctggac gcagcttcgg ggggcctgct 1550 ggctgccttc ccccgctgcgtggtccgagt gcctgtggct cgctgccagc 1600 agtactcggg gtgtatgaag aactgtatcggcagtcagga cccctactgc 1650 gggtgggccc ccgacggctc ctgcatcttc ctcagcccgggcaccagagc 1700 cgcctttgag caggacgtgt ccggggccag cacctcaggc ttaggggact1750 gcacaggact cctgcgggcc agcctctccg aggaccgcgc ggggctggtg 1800tcggtgaacc tgctggtaac gtcgtcggtg gcggccttcg tggtgggagc 1850 cgtggtgtccggcttcagcg tgggctggtt cgtgggcctc cgtgagcggc 1900 gggagctggc ccggcgcaaggacaaggagg ccatcctggc gcacggggcg 1950 ggcgaggcgg tgctgagcgt cagccgcctgggcgagcgca gggcgcaggg 2000 tcccgggggc cggggcggag gcggtggcgg tggcgccggggttcccccgg 2050 aggccctgct ggcgcccctg atgcagaacg gctgggccaa ggccacgctg2100 ctgcagggcg ggccccacga cctggactcg gggctgctgc ccacgcccga 2150gcagacgccg ctgccgcaga agcgcctgcc cactccgcac ccgcaccccc 2200 acgccctgggcccccgcgcc tgggaccacg gccaccccct gctcccggcc 2250 tccgcttcat cctccctcctgctgctggcg cccgcccggg cccccgagca 2300 gccccccgcg cctggggagc cgacccccgacggccgcctc tatgctgccc 2350 ggcccggccg cgcctcccac ggcgacttcc cgctcaccccccacgccagc 2400 ccggaccgcc ggcgggtggt gtccgcgccc acgggcccct tggacccagc2450 ctcagccgcc gatggcctcc cgcggccctg gagcccgccc ccgacgggca 2500gcctgaggag gccactgggc ccccacgccc ctccggccgc caccctgcgc 2550 cgcacccacacgttcaacag cggcgaggcc cggcctgggg accgccaccg 2600 cggctgccac gcccggccgggcacagactt ggcccacctc ctcccctatg 2650 ggggggcgga caggactgcg ccccccgtgccctaggccgg gggccccccg 2700 atgccttggc agtgccagcc acgggaacca ggagcgagagacggtgccag 2750 aacgccgggg cccggggcaa ctccgagtgg gtgctcaagt cccccccgcg2800 acccacccgc ggagtggggg gccccctccg ccacaaggaa gcacaaccag 2850ctcgccctcc ccctacccgg ggccgcagga cgctgagacg gtttgggggt 2900 gggtgggcgggaggactttg ctatggattt gaggttgacc ttatgcgcgt 2950 aggttttggt ttttttttgcagttttggtt tcttttgcgg ttttctaacc 3000 aattgcacaa ctccgttctc ggggtggcggcaggcagggg aggcttggac 3050 gccggtgggg aatggggggc cacagctgca gacctaagccctcccccacc 3100 cctggaaagg tccctcccca acccaggccc ctggcgtgtg tgggtgtgcg3150 tgcgtgtgcg tgccgtgttc gtgtgcaagg ggccggggag gtgggcgtgt 3200gtgtgcgtgc cagcgaaggc tgctgtgggc gtgtgtgtca agtgggccac 3250 gcgtgcagggtgtgtgtcca cgagcgacga tcgtggtggc cccagcggcc 3300 tgggcgttgg ctgagccgacgctggggctt ccagaaggcc cgggggtctc 3350 cgaggtgccg gttaggagtt tgaaccccccccactctgca gagggaagcg 3400 gggacaatgc cggggtttca ggcaggagac acgaggagggcctgcccgga 3450 agtcacatcg gcagcagctg tctaaagggc ttgggggcct ggggggcggc3500 gaaggtgggt ggggcccctc tgtaaatacg gccccagggt ggtgagagag 3550tcccatgcca cccgtcccct tgtgacctcc cccctatgac ctccagctga 3600 ccatgcatgccacgtggctg gctgggtcct ctgccctctt tggagtttgc 3650 ctcccccagc cccctccccatcaataaaac tctgtttaca accaaaaaaa 3700 aaaaaaaaaa aaaaaaaaaa a 3721 35888 PRT Homo Sapien 35 Met Gln Thr Pro Arg Ala Ser Pro Pro Arg Pro AlaLeu Leu Leu 1 5 10 15 Leu Leu Leu Leu Leu Gly Gly Ala His Gly Leu PhePro Glu Glu 20 25 30 Pro Pro Pro Leu Ser Val Ala Pro Arg Asp Tyr Leu AsnHis Tyr 35 40 45 Pro Val Phe Val Gly Ser Gly Pro Gly Arg Leu Thr Pro AlaGlu 50 55 60 Gly Ala Asp Asp Leu Asn Ile Gln Arg Val Leu Arg Val Asn Arg65 70 75 Thr Leu Phe Ile Gly Asp Arg Asp Asn Leu Tyr Arg Val Glu Leu 8085 90 Glu Pro Pro Thr Ser Thr Glu Leu Arg Tyr Gln Arg Lys Leu Thr 95 100105 Trp Arg Ser Asn Pro Ser Asp Ile Asn Val Cys Arg Met Lys Gly 110 115120 Lys Gln Glu Gly Glu Cys Arg Asn Phe Val Lys Val Leu Leu Leu 125 130135 Arg Asp Glu Ser Thr Leu Phe Val Cys Gly Ser Asn Ala Phe Asn 140 145150 Pro Val Cys Ala Asn Tyr Ser Ile Asp Thr Leu Gln Pro Val Gly 155 160165 Asp Asn Ile Ser Gly Met Ala Arg Cys Pro Tyr Asp Pro Lys His 170 175180 Ala Asn Val Ala Leu Phe Ser Asp Gly Met Leu Phe Thr Ala Thr 185 190195 Val Thr Asp Phe Leu Ala Ile Asp Ala Val Ile Tyr Arg Ser Leu 200 205210 Gly Asp Arg Pro Thr Leu Arg Thr Val Lys His Asp Ser Lys Trp 215 220225 Phe Lys Glu Pro Tyr Phe Val His Ala Val Glu Trp Gly Ser His 230 235240 Val Tyr Phe Phe Phe Arg Glu Ile Ala Met Glu Phe Asn Tyr Leu 245 250255 Glu Lys Val Val Val Ser Arg Val Ala Arg Val Cys Lys Asn Asp 260 265270 Val Gly Gly Ser Pro Arg Val Leu Glu Lys Gln Trp Thr Ser Phe 275 280285 Leu Lys Ala Arg Leu Asn Cys Ser Val Pro Gly Asp Ser His Phe 290 295300 Tyr Phe Asn Val Leu Gln Ala Val Thr Gly Val Val Ser Leu Gly 305 310315 Gly Arg Pro Val Val Leu Ala Val Phe Ser Thr Pro Ser Asn Ser 320 325330 Ile Pro Gly Ser Ala Val Cys Ala Phe Asp Leu Thr Gln Val Ala 335 340345 Ala Val Phe Glu Gly Arg Phe Arg Glu Gln Lys Ser Pro Glu Ser 350 355360 Ile Trp Thr Pro Val Pro Glu Asp Gln Val Pro Arg Pro Arg Pro 365 370375 Gly Cys Cys Ala Ala Pro Gly Met Gln Tyr Asn Ala Ser Ser Ala 380 385390 Leu Pro Asp Asp Ile Leu Asn Phe Val Lys Thr His Pro Leu Met 395 400405 Asp Glu Ala Val Pro Ser Leu Gly His Ala Pro Trp Ile Leu Arg 410 415420 Thr Leu Met Arg His Gln Leu Thr Arg Val Ala Val Asp Val Gly 425 430435 Ala Gly Pro Trp Gly Asn Gln Thr Val Val Phe Leu Gly Ser Glu 440 445450 Ala Gly Thr Val Leu Lys Phe Leu Val Arg Pro Asn Ala Ser Thr 455 460465 Ser Gly Thr Ser Gly Leu Ser Val Phe Leu Glu Glu Phe Glu Thr 470 475480 Tyr Arg Pro Asp Arg Cys Gly Arg Pro Gly Gly Gly Glu Thr Gly 485 490495 Gln Arg Leu Leu Ser Leu Glu Leu Asp Ala Ala Ser Gly Gly Leu 500 505510 Leu Ala Ala Phe Pro Arg Cys Val Val Arg Val Pro Val Ala Arg 515 520525 Cys Gln Gln Tyr Ser Gly Cys Met Lys Asn Cys Ile Gly Ser Gln 530 535540 Asp Pro Tyr Cys Gly Trp Ala Pro Asp Gly Ser Cys Ile Phe Leu 545 550555 Ser Pro Gly Thr Arg Ala Ala Phe Glu Gln Asp Val Ser Gly Ala 560 565570 Ser Thr Ser Gly Leu Gly Asp Cys Thr Gly Leu Leu Arg Ala Ser 575 580585 Leu Ser Glu Asp Arg Ala Gly Leu Val Ser Val Asn Leu Leu Val 590 595600 Thr Ser Ser Val Ala Ala Phe Val Val Gly Ala Val Val Ser Gly 605 610615 Phe Ser Val Gly Trp Phe Val Gly Leu Arg Glu Arg Arg Glu Leu 620 625630 Ala Arg Arg Lys Asp Lys Glu Ala Ile Leu Ala His Gly Ala Gly 635 640645 Glu Ala Val Leu Ser Val Ser Arg Leu Gly Glu Arg Arg Ala Gln 650 655660 Gly Pro Gly Gly Arg Gly Gly Gly Gly Gly Gly Gly Ala Gly Val 665 670675 Pro Pro Glu Ala Leu Leu Ala Pro Leu Met Gln Asn Gly Trp Ala 680 685690 Lys Ala Thr Leu Leu Gln Gly Gly Pro His Asp Leu Asp Ser Gly 695 700705 Leu Leu Pro Thr Pro Glu Gln Thr Pro Leu Pro Gln Lys Arg Leu 710 715720 Pro Thr Pro His Pro His Pro His Ala Leu Gly Pro Arg Ala Trp 725 730735 Asp His Gly His Pro Leu Leu Pro Ala Ser Ala Ser Ser Ser Leu 740 745750 Leu Leu Leu Ala Pro Ala Arg Ala Pro Glu Gln Pro Pro Ala Pro 755 760765 Gly Glu Pro Thr Pro Asp Gly Arg Leu Tyr Ala Ala Arg Pro Gly 770 775780 Arg Ala Ser His Gly Asp Phe Pro Leu Thr Pro His Ala Ser Pro 785 790795 Asp Arg Arg Arg Val Val Ser Ala Pro Thr Gly Pro Leu Asp Pro 800 805810 Ala Ser Ala Ala Asp Gly Leu Pro Arg Pro Trp Ser Pro Pro Pro 815 820825 Thr Gly Ser Leu Arg Arg Pro Leu Gly Pro His Ala Pro Pro Ala 830 835840 Ala Thr Leu Arg Arg Thr His Thr Phe Asn Ser Gly Glu Ala Arg 845 850855 Pro Gly Asp Arg His Arg Gly Cys His Ala Arg Pro Gly Thr Asp 860 865870 Leu Ala His Leu Leu Pro Tyr Gly Gly Ala Asp Arg Thr Ala Pro 875 880885 Pro Val Pro 36 21 DNA Artificial Sequence Synthetic oligonucleotideprobe 36 gaggacctac cggccggaca g 21 37 24 DNA Artificial SequenceSynthetic oligonucleotide probe 37 atacaccccg agtactgctg gcag 24 38 42DNA Artificial Sequence Synthetic oligonucleotide probe 38 agacagggcagcggctgctg agcttggagc tggacgcagc tt 42 39 2014 DNA Homo Sapien 39agcaactcaa gttcatcatt gtcctgagag agaggagcag cgcggttctc 50 ggccgggacagcagaacgcc aggggaccct cacctgggcg cgccggggca 100 cgggctttga ttgtcctggggtcgcggaga cccgcgcgcc tgccctgcac 150 gccgggcggc aacctttgca gtcgcgttggctgctgcgat cggccggcgg 200 gtccctgccg aaggctcggc tgcttctgtc cacctcttacacttcttcat 250 ttatcggtgg atcatttcga gagtccgtct tgtaaatgtt tggcactttg300 ctactttatt gcttctttct ggcgacagtt ccagcactcg ccgagaccgg 350cggagaaagg cagctgagcc cggagaagag cgaaatatgg ggacccgggc 400 taaaagcagacgtcgtcctt cccgcccgct atttctatat tcaggcagtg 450 gatacatcag ggaataaattcacatcttct ccaggcgaaa aggtcttcca 500 ggtgaaagtc tcagcaccag aggagcaattcactagagtt ggagtccagg 550 ttttagaccg aaaagatggg tccttcatag taagatacagaatgtatgca 600 agctacaaaa atctgaaggt ggaaattaaa ttccaagggc aacatgtggc650 caaatcccca tatattttaa aagggccggt ttaccatgag aactgtgact 700gtcctctgca agatagtgca gcctggctac gggagatgaa ctgccctgaa 750 accattgctcagattcagag agatctggca catttccctg ctgtggatcc 800 agaaaagatt gcagtagaaatcccaaaaag atttggacag aggcagagcc 850 tatgtcacta caccttaaag gataacaaggtttatatcaa gactcatggt 900 gaacatgtag gttttagaat tttcatggat gccatactactttctttgac 950 tagaaaggtg aagatgccag atgtggagct ctttgttaat ttgggagact1000 ggcctttgga aaaaaagaaa tccaattcaa acatccatcc gatcttttcc 1050tggtgtggct ccacagattc caaggatatc gtgatgccta cgtacgattt 1100 gactgattctgttctggaaa ccatgggccg ggtaagtctg gatatgatgt 1150 ccgtgcaagc taacacgggtcctccctggg aaagcaaaaa ttccactgcc 1200 gtctggagag ggcgagacag ccgcaaagagagactcgagc tggttaaact 1250 cagtagaaaa cacccagaac tcatagacgc tgctttcaccaactttttct 1300 tctttaaaca cgatgaaaac ctgtatggtc ccattgtgaa acatatttca1350 ttttttgatt tcttcaagca taagtatcaa ataaatatcg atggcactgt 1400agcagcttat cgcctgccat atttgctagt tggtgacagt gttgtgctga 1450 agcaggattccatctactat gaacattttt acaatgagct gcagccctgg 1500 aaacactaca ttccagttaagagcaacctg agcgatctgc tagaaaaact 1550 taaatgggcg aaagatcacg atgaagaggccaaaaagata gcaaaagcag 1600 gacaagaatt tgcaagaaat aatctcatgg gcgatgacatattctgttat 1650 tatttcaaac ttttccagga atatgccaat ttacaagtga gtgagcccca1700 aatccgagag ggcatgaaaa gggtagaacc acagactgag gacgacctct 1750tcccttgtac ttgccatagg aaaaagacca aagatgaact ctgatatgca 1800 aaataacttctattagaata atggtgctct gaagactctt cttaactaaa 1850 aagaagaatt tttttaagtattaattccat ggacaatata aaatctgtgt 1900 gattgtttgc agtatgaaga cacatttctacttatgcagt attctcatga 1950 ctgtacttta aagtacattt ttagaatttt ataataaaaccacctttatt 2000 ttaaaggaaa aaaa 2014 40 502 PRT Homo Sapien 40 Met PheGly Thr Leu Leu Leu Tyr Cys Phe Phe Leu Ala Thr Val 1 5 10 15 Pro AlaLeu Ala Glu Thr Gly Gly Glu Arg Gln Leu Ser Pro Glu 20 25 30 Lys Ser GluIle Trp Gly Pro Gly Leu Lys Ala Asp Val Val Leu 35 40 45 Pro Ala Arg TyrPhe Tyr Ile Gln Ala Val Asp Thr Ser Gly Asn 50 55 60 Lys Phe Thr Ser SerPro Gly Glu Lys Val Phe Gln Val Lys Val 65 70 75 Ser Ala Pro Glu Glu GlnPhe Thr Arg Val Gly Val Gln Val Leu 80 85 90 Asp Arg Lys Asp Gly Ser PheIle Val Arg Tyr Arg Met Tyr Ala 95 100 105 Ser Tyr Lys Asn Leu Lys ValGlu Ile Lys Phe Gln Gly Gln His 110 115 120 Val Ala Lys Ser Pro Tyr IleLeu Lys Gly Pro Val Tyr His Glu 125 130 135 Asn Cys Asp Cys Pro Leu GlnAsp Ser Ala Ala Trp Leu Arg Glu 140 145 150 Met Asn Cys Pro Glu Thr IleAla Gln Ile Gln Arg Asp Leu Ala 155 160 165 His Phe Pro Ala Val Asp ProGlu Lys Ile Ala Val Glu Ile Pro 170 175 180 Lys Arg Phe Gly Gln Arg GlnSer Leu Cys His Tyr Thr Leu Lys 185 190 195 Asp Asn Lys Val Tyr Ile LysThr His Gly Glu His Val Gly Phe 200 205 210 Arg Ile Phe Met Asp Ala IleLeu Leu Ser Leu Thr Arg Lys Val 215 220 225 Lys Met Pro Asp Val Glu LeuPhe Val Asn Leu Gly Asp Trp Pro 230 235 240 Leu Glu Lys Lys Lys Ser AsnSer Asn Ile His Pro Ile Phe Ser 245 250 255 Trp Cys Gly Ser Thr Asp SerLys Asp Ile Val Met Pro Thr Tyr 260 265 270 Asp Leu Thr Asp Ser Val LeuGlu Thr Met Gly Arg Val Ser Leu 275 280 285 Asp Met Met Ser Val Gln AlaAsn Thr Gly Pro Pro Trp Glu Ser 290 295 300 Lys Asn Ser Thr Ala Val TrpArg Gly Arg Asp Ser Arg Lys Glu 305 310 315 Arg Leu Glu Leu Val Lys LeuSer Arg Lys His Pro Glu Leu Ile 320 325 330 Asp Ala Ala Phe Thr Asn PhePhe Phe Phe Lys His Asp Glu Asn 335 340 345 Leu Tyr Gly Pro Ile Val LysHis Ile Ser Phe Phe Asp Phe Phe 350 355 360 Lys His Lys Tyr Gln Ile AsnIle Asp Gly Thr Val Ala Ala Tyr 365 370 375 Arg Leu Pro Tyr Leu Leu ValGly Asp Ser Val Val Leu Lys Gln 380 385 390 Asp Ser Ile Tyr Tyr Glu HisPhe Tyr Asn Glu Leu Gln Pro Trp 395 400 405 Lys His Tyr Ile Pro Val LysSer Asn Leu Ser Asp Leu Leu Glu 410 415 420 Lys Leu Lys Trp Ala Lys AspHis Asp Glu Glu Ala Lys Lys Ile 425 430 435 Ala Lys Ala Gly Gln Glu PheAla Arg Asn Asn Leu Met Gly Asp 440 445 450 Asp Ile Phe Cys Tyr Tyr PheLys Leu Phe Gln Glu Tyr Ala Asn 455 460 465 Leu Gln Val Ser Glu Pro GlnIle Arg Glu Gly Met Lys Arg Val 470 475 480 Glu Pro Gln Thr Glu Asp AspLeu Phe Pro Cys Thr Cys His Arg 485 490 495 Lys Lys Thr Lys Asp Glu Leu500 41 26 DNA Artificial Sequence Synthetic oligonucleotide probe 41gaaggtggaa attaaattcc aagggc 26 42 24 DNA Artificial Sequence Syntheticoligonucleotide probe 42 cgataagctg ctacagtgcc atcg 24 43 40 DNAArtificial Sequence Synthetic oligonucleotide probe 43 gtgactgtcctctgcaagat agtgcagcct ggctacggga 40 44 2395 DNA Homo Sapien 44cctggagccg gaagcgcggc tgcagcaggg cgaggctcca ggtggggtcg 50 gttccgcatccagcctagcg tgtccacgat gcggctgggc tccgggactt 100 tcgctacctg ttgcgtagcgatcgaggtgc tagggatcgc ggtcttcctt 150 cggggattct tcccggctcc cgttcgttcctctgccagag cggaacacgg 200 agcggagccc ccagcgcccg aaccctcggc tggagccagttctaactgga 250 ccacgctgcc accacctctc ttcagtaaag ttgttattgt tctgatagat300 gccttgagag atgattttgt gtttgggtca aagggtgtga aatttatgcc 350ctacacaact taccttgtgg aaaaaggagc atctcacagt tttgtggctg 400 aagcaaagccacctacagtt actatgcctc gaatcaaggc attgatgacg 450 gggagccttc ctggctttgtcgacgtcatc aggaacctca attctcctgc 500 actgctggaa gacagtgtga taagacaagcaaaagcagct ggaaaaagaa 550 tagtctttta tggagatgaa acctgggtta aattattcccaaagcatttt 600 gtggaatatg atggaacaac ctcatttttc gtgtcagatt acacagaggt650 ggataataat gtcacgaggc atttggataa agtattaaaa agaggagatt 700gggacatatt aatcctccac tacctggggc tggaccacat tggccacatt 750 tcagggcccaacagccccct gattgggcag aagctgagcg agatggacag 800 cgtgctgatg aagatccacacctcactgca gtcgaaggag agagagacgc 850 ctttacccaa tttgctggtt ctttgtggtgaccatggcat gtctgaaaca 900 ggaagtcacg gggcctcctc caccgaggag gtgaatacacctctgatttt 950 aatcagttct gcgtttgaaa ggaaacccgg tgatatccga catccaaagc1000 acgtccaata gacggatgtg gctgcgacac tggcgatagc acttggctta 1050ccgattccaa aagacagtgt agggagcctc ctattcccag ttgtggaagg 1100 aagaccaatgagagagcagt tgagattttt acatttgaat acagtgcagc 1150 ttagtaaact gttgcaagagaatgtgccgt catatgaaaa agatcctggg 1200 tttgagcagt ttaaaatgtc agaaagattgcatgggaact ggatcagact 1250 gtacttggag gaaaagcatt cagaagtcct attcaacctgggctccaagg 1300 ttctcaggca gtacctggat gctctgaaga cgctgagctt gtccctgagt1350 gcacaagtgg cccagttctc accctgctcc tgctcagcgt cccacaggca 1400ctgcacagaa aggctgagct ggaagtccca ctgtcatctc ctgggttttc 1450 tctgctcttttatttggtga tcctggttct ttcggccgtt cacgtcattg 1500 tgtgcacctc agctgaaagttcgtgctact tctgtggcct ctcgtggctg 1550 gcggcaggct gcctttcgtt taccagactctggttgaaca cctggtgtgt 1600 gccaagtgct ggcagtgccc tggacagggg gcctcagggaaggacgtgga 1650 gcagccttat cccaggcctc tgggtgtccc gacacaggtg ttcacatctg1700 tgctgtcagg tcagatgcct cagttcttgg aaagctaggt tcctgcgact 1750gttaccaagg tgattgtaaa gagctggcgg tcacagagga acaagccccc 1800 cagctgagggggtgtgtgaa tcggacagcc tcccagcaga ggtgtgggag 1850 ctgcagctga gggaagaagagacaatcggc ctggacactc aggagggtca 1900 aaaggagact tggtcgcacc actcatcctgccacccccag aatgcatcct 1950 gcctcatcag gtccagattt ctttccaagg cggacgttttctgttggaat 2000 tcttagtcct tggcctcgga caccttcatt cgttagctgg ggagtggtgg2050 tgaggcagtg aagaagaggc ggatggtcac actcagatcc acagagccca 2100ggatcaaggg acccactgca gtggcagcag gactgttggg cccccacccc 2150 aaccctgcacagccctcatc ccctcttggc ttgagccgtc agaggccctg 2200 tgctgagtgt ctgaccgagacactcacagc tttgtcatca gggcacaggc 2250 ttcctcggag ccaggatgat ctgtgccacgcttgcacctc gggcccatct 2300 gggctcatgc tctctctcct gctattgaat tagtacctagctgcacacag 2350 tatgtagtta ccaaaagaat aaacggcaat aattgagaaa aaaaa 239545 310 PRT Homo Sapien 45 Met Arg Leu Gly Ser Gly Thr Phe Ala Thr CysCys Val Ala Ile 1 5 10 15 Glu Val Leu Gly Ile Ala Val Phe Leu Arg GlyPhe Phe Pro Ala 20 25 30 Pro Val Arg Ser Ser Ala Arg Ala Glu His Gly AlaGlu Pro Pro 35 40 45 Ala Pro Glu Pro Ser Ala Gly Ala Ser Ser Asn Trp ThrThr Leu 50 55 60 Pro Pro Pro Leu Phe Ser Lys Val Val Ile Val Leu Ile AspAla 65 70 75 Leu Arg Asp Asp Phe Val Phe Gly Ser Lys Gly Val Lys Phe Met80 85 90 Pro Tyr Thr Thr Tyr Leu Val Glu Lys Gly Ala Ser His Ser Phe 95100 105 Val Ala Glu Ala Lys Pro Pro Thr Val Thr Met Pro Arg Ile Lys 110115 120 Ala Leu Met Thr Gly Ser Leu Pro Gly Phe Val Asp Val Ile Arg 125130 135 Asn Leu Asn Ser Pro Ala Leu Leu Glu Asp Ser Val Ile Arg Gln 140145 150 Ala Lys Ala Ala Gly Lys Arg Ile Val Phe Tyr Gly Asp Glu Thr 155160 165 Trp Val Lys Leu Phe Pro Lys His Phe Val Glu Tyr Asp Gly Thr 170175 180 Thr Ser Phe Phe Val Ser Asp Tyr Thr Glu Val Asp Asn Asn Val 185190 195 Thr Arg His Leu Asp Lys Val Leu Lys Arg Gly Asp Trp Asp Ile 200205 210 Leu Ile Leu His Tyr Leu Gly Leu Asp His Ile Gly His Ile Ser 215220 225 Gly Pro Asn Ser Pro Leu Ile Gly Gln Lys Leu Ser Glu Met Asp 230235 240 Ser Val Leu Met Lys Ile His Thr Ser Leu Gln Ser Lys Glu Arg 245250 255 Glu Thr Pro Leu Pro Asn Leu Leu Val Leu Cys Gly Asp His Gly 260265 270 Met Ser Glu Thr Gly Ser His Gly Ala Ser Ser Thr Glu Glu Val 275280 285 Asn Thr Pro Leu Ile Leu Ile Ser Ser Ala Phe Glu Arg Lys Pro 290295 300 Gly Asp Ile Arg His Pro Lys His Val Gln 305 310 46 22 DNAArtificial Sequence Synthetic oligonucleotide probe 46 cgggactttcgctacctgtt gc 22 47 26 DNA Artificial Sequence Synthetic oligonucleotideprobe 47 catcatattc cacaaaatgc tttggg 26 48 38 DNA Artificial SequenceSynthetic oligonucleotide probe 48 ccttcgggga ttcttcccgg ctcccgttcgttcctctg 38 49 918 DNA Homo Sapien 49 agccaggcag cacatcacag cgggaggagctgtcccaggt ggcccagctc 50 agcaatggca atgggggtcc ccagagtcat tctgctctgcctctttgggg 100 ctgcgctctg cctgacaggg tcccaagccc tgcagtgcta cagctttgag150 cacacctact ttggcccctt tgacctcagg gccatgaagc tgcccagcat 200ctcctgtcct catgagtgct ttgaggctat cctgtctctg gacaccgggt 250 atcgcgcgccggtgaccctg gtgcggaagg gctgctggac cgggcctcct 300 gcgggccaga cgcaatcgaacccggacgcg ctgccgccag actactcggt 350 ggtgcgcggc tgcacaactg acaaatgcaacgcccacctc atgactcatg 400 acgccctccc caacctgagc caagcacccg acccgccgacgctcagcggc 450 gccgagtgct acgcctgtat cggggtccac caggatgact gcgctatcgg500 caggtcccga cgagtccagt gtcaccagga ccagaccgcc tgcttccagg 550gcagtggcag aatgacagtt ggcaatttct cagtccctgt gtacatcaga 600 acctgccaccggccctcctg caccaccgag ggcaccacca gcccctggac 650 agccatcgac ctccagggctcctgctgtga ggggtacctc tgcaacagga 700 aatccatgac ccagcccttc accagtgcttcagccaccac ccctccccga 750 gcactacagg tcctggccct gctcctccca gtcctcctgctggtggggct 800 ctcagcatag accgcccctc caggatgctg gggacagggc tcacacacct850 cattcttgct gcttcagccc ctatcacata gctcactgga aaatgatgtt 900aaagtaagaa ttgcaaaa 918 50 251 PRT Homo Sapien 50 Met Ala Met Gly ValPro Arg Val Ile Leu Leu Cys Leu Phe Gly 1 5 10 15 Ala Ala Leu Cys LeuThr Gly Ser Gln Ala Leu Gln Cys Tyr Ser 20 25 30 Phe Glu His Thr Tyr PheGly Pro Phe Asp Leu Arg Ala Met Lys 35 40 45 Leu Pro Ser Ile Ser Cys ProHis Glu Cys Phe Glu Ala Ile Leu 50 55 60 Ser Leu Asp Thr Gly Tyr Arg AlaPro Val Thr Leu Val Arg Lys 65 70 75 Gly Cys Trp Thr Gly Pro Pro Ala GlyGln Thr Gln Ser Asn Pro 80 85 90 Asp Ala Leu Pro Pro Asp Tyr Ser Val ValArg Gly Cys Thr Thr 95 100 105 Asp Lys Cys Asn Ala His Leu Met Thr HisAsp Ala Leu Pro Asn 110 115 120 Leu Ser Gln Ala Pro Asp Pro Pro Thr LeuSer Gly Ala Glu Cys 125 130 135 Tyr Ala Cys Ile Gly Val His Gln Asp AspCys Ala Ile Gly Arg 140 145 150 Ser Arg Arg Val Gln Cys His Gln Asp GlnThr Ala Cys Phe Gln 155 160 165 Gly Ser Gly Arg Met Thr Val Gly Asn PheSer Val Pro Val Tyr 170 175 180 Ile Arg Thr Cys His Arg Pro Ser Cys ThrThr Glu Gly Thr Thr 185 190 195 Ser Pro Trp Thr Ala Ile Asp Leu Gln GlySer Cys Cys Glu Gly 200 205 210 Tyr Leu Cys Asn Arg Lys Ser Met Thr GlnPro Phe Thr Ser Ala 215 220 225 Ser Ala Thr Thr Pro Pro Arg Ala Leu GlnVal Leu Ala Leu Leu 230 235 240 Leu Pro Val Leu Leu Leu Val Gly Leu SerAla 245 250 51 3288 DNA Homo Sapien 51 cccacgcgtc cgggacagat gaacttaaaagagaagcttt agctgccaaa 50 gattgggaaa gggaaaggac aaaaaagacc cctgggctacacggcgtagg 100 tgcagggttt cctactgctg ttcttttatg ctgggagctg tggctgtaac150 caactaggaa ataacgtatg cagcagctat ggctgtcaga gagttgtgct 200tcccaagaca aaggcaagtc ctgtttcttt ttcttttttg gggagtgtcc 250 ttggcaggttctgggtttgg acgttattcg gtgactgagg aaacagagaa 300 aggatccttt gtggtcaatctggcaaagga tctgggacta gcagaggggg 350 agctggctgc aaggggaacc agggtggtttccgatgataa caaacaatac 400 ctgctcctgg attcacatac cgggaatttg ctcacaaatgagaaactgga 450 ccgagagaag ctgtgtggcc ctaaagagcc ctgtatgctg tatttccaaa500 ttttaatgga tgatcccttt cagatttacc gggctgagct gagagtcagg 550gatataaatg atcacgcgcc agtatttcag gacaaagaaa cagtcttaaa 600 aatatcagaaaatacagctg aagggacagc atttagacta gaaagagcac 650 aggatccaga tggaggacttaacggtatcc aaaactacac gatcagcccc 700 aactcttttt tccatattaa cattagtggcggtgatgaag gcatgatata 750 tccagagcta gtgttggaca aagcactgga tcgggaggagcagggagagc 800 tcagcttaac cctcacagcg ctggatggtg ggtctccatc caggtctggg850 acctctactg tacgcatcgt tgtcttggac gtcaatgaca atgccccaca 900gtttgcccag gctctgtatg agacccaggc tccagaaaac agccccattg 950 ggttccttattgttaaggta tgggcagaag atgtagactc tggagtcaac 1000 gcggaagtat cctattcattttttgatgcc tcagaaaata ttcgaacgac 1050 ctttcaaatc aatccttttt ctggggaaatctttctcaga gaattgcttg 1100 attatgagtt agtaaattct tacaaaataa atatacaggcaatggacggt 1150 ggaggccttt ctgcaagatg tagggtttta gtggaagtat tggacaccaa1200 tgacaatccc cctgaactga tcgtatcatc attttccaac tctgttgctg 1250agaattctcc tgagacgccg ctggctgttt ttaagattaa tgacagagac 1300 tctggagaaaatggaaagat ggtttgctac attcaagaga atctgccatt 1350 cctactaaaa ccttctgtggagaattttta catcctaatt acagaaggcg 1400 cgctggacag agagatcaga gccgagtacaacatcactat caccgtcact 1450 gacttgggga cacccaggct gaaaaccgag cacaacataacggtcctggt 1500 ctccgacgtc aatgacaacg cccccgcctt cacccaaacc tcctacaccc1550 tgttcgtccg cgagaacaac agccccgccc tgcacatcgg cagcgtcagc 1600gccacagaca gagactcggg caccaacgcc caggtcacct actcgctgct 1650 gccgccccaagacccgcacc tgcccctcgc ctccctggtc tccatcaacg 1700 cggacaacgg ccacctgttcgccctcaggt cgctggacta cgaggccctg 1750 caggctttcg agttccgcgt gggcgccacagaccgcggct cccccgcgct 1800 gagcagagag gcgctggtgc gcgtgctggt gctggacgccaacgacaact 1850 cgcccttcgt gctgtacccg ctgcagaacg gctccgcgcc ctgcaccgag1900 ctggtgcccc gggcggccga gccgggctac ctggtgacca aggtggtggc 1950ggtggacggc gactcgggcc agaacgcctg gctgtcgtac cagctgctca 2000 aggccacggagcccgggctg ttcggtgtgt gggcgcacaa tggggaggtg 2050 cgcaccgcca ggctgctgagcgagcgcgac gcagccaagc acaggctcgt 2100 ggtgcttgtc aaggacaatg gcgagcctcctcgctcggcc accgccacgc 2150 tgcacttgct cctggtggac ggcttctccc agccctacctgcctctcccg 2200 gaggcggccc cggcccaggc ccaggccgag gccgacttgc tcaccgtcta2250 cctggtggtg gcgttggcct cggtgtcttc gctcttcctc ctctcggtgc 2300tcctgttcgt ggcggtgcgg ctgtgcagga ggagcagggc ggcctcggtg 2350 ggtcgctgctcggtgcccga gggtcctttt ccagggcatc tggtggacgt 2400 gaggggcgct gagaccctgtcccagagcta ccagtatgag gtgtgtctga 2450 cgggaggccc cgggaccagt gagttcaagttcttgaaacc agttatttcg 2500 gatattcagg cacagggccc tgggaggaag ggtgaagaaaattccacctt 2550 ccgaaatagc tttggattta atattcagta aagtctgttt ttagtttcat2600 atacttttgg tgtgttacat agccatgttt ctattagttt acttttaaat 2650ctcaaattta agttattatg caacttcaag cattattttc aagtagtata 2700 cccctgtggttttacaatgt ttcatcattt ttttgcatta ataacaactg 2750 ggtttaattt aatgagtatttttttctaaa tgatagtgtt aaggttttaa 2800 ttctttccaa ctgcccaagg aattaattactattatatct cattacagaa 2850 atctgaggtt ttgattcatt tcagagcttg catctcatgattctaatcac 2900 ttctgtctat agtgtacttg ctctatttaa gaaggcatat ctacatttcc2950 aaactcattc taacattcta tatattcgtg tttgaaaacc atgtcattta 3000tttctacatc atgtatttaa aaagaaatat ttctctacta ctatgctcat 3050 gacaaaatgaaacaaagcat attgtgagca atactgaaca tcaataatac 3100 ccttagttta tatacttattattttatctt taagcatgct acttttactt 3150 ggccaatatt ttcttatgtt aacttttgctgatgtataaa acagactatg 3200 ccttataatt gaaataaaat tataatctgc ctgaaaatgaataaaaataa 3250 aacattttga aatgtgaaaa aaaaaaaaaa aaaaaaaa 3288 52 800PRT Homo Sapien 52 Met Ala Val Arg Glu Leu Cys Phe Pro Arg Gln Arg GlnVal Leu 1 5 10 15 Phe Leu Phe Leu Phe Trp Gly Val Ser Leu Ala Gly SerGly Phe 20 25 30 Gly Arg Tyr Ser Val Thr Glu Glu Thr Glu Lys Gly Ser PheVal 35 40 45 Val Asn Leu Ala Lys Asp Leu Gly Leu Ala Glu Gly Glu Leu Ala50 55 60 Ala Arg Gly Thr Arg Val Val Ser Asp Asp Asn Lys Gln Tyr Leu 6570 75 Leu Leu Asp Ser His Thr Gly Asn Leu Leu Thr Asn Glu Lys Leu 80 8590 Asp Arg Glu Lys Leu Cys Gly Pro Lys Glu Pro Cys Met Leu Tyr 95 100105 Phe Gln Ile Leu Met Asp Asp Pro Phe Gln Ile Tyr Arg Ala Glu 110 115120 Leu Arg Val Arg Asp Ile Asn Asp His Ala Pro Val Phe Gln Asp 125 130135 Lys Glu Thr Val Leu Lys Ile Ser Glu Asn Thr Ala Glu Gly Thr 140 145150 Ala Phe Arg Leu Glu Arg Ala Gln Asp Pro Asp Gly Gly Leu Asn 155 160165 Gly Ile Gln Asn Tyr Thr Ile Ser Pro Asn Ser Phe Phe His Ile 170 175180 Asn Ile Ser Gly Gly Asp Glu Gly Met Ile Tyr Pro Glu Leu Val 185 190195 Leu Asp Lys Ala Leu Asp Arg Glu Glu Gln Gly Glu Leu Ser Leu 200 205210 Thr Leu Thr Ala Leu Asp Gly Gly Ser Pro Ser Arg Ser Gly Thr 215 220225 Ser Thr Val Arg Ile Val Val Leu Asp Val Asn Asp Asn Ala Pro 230 235240 Gln Phe Ala Gln Ala Leu Tyr Glu Thr Gln Ala Pro Glu Asn Ser 245 250255 Pro Ile Gly Phe Leu Ile Val Lys Val Trp Ala Glu Asp Val Asp 260 265270 Ser Gly Val Asn Ala Glu Val Ser Tyr Ser Phe Phe Asp Ala Ser 275 280285 Glu Asn Ile Arg Thr Thr Phe Gln Ile Asn Pro Phe Ser Gly Glu 290 295300 Ile Phe Leu Arg Glu Leu Leu Asp Tyr Glu Leu Val Asn Ser Tyr 305 310315 Lys Ile Asn Ile Gln Ala Met Asp Gly Gly Gly Leu Ser Ala Arg 320 325330 Cys Arg Val Leu Val Glu Val Leu Asp Thr Asn Asp Asn Pro Pro 335 340345 Glu Leu Ile Val Ser Ser Phe Ser Asn Ser Val Ala Glu Asn Ser 350 355360 Pro Glu Thr Pro Leu Ala Val Phe Lys Ile Asn Asp Arg Asp Ser 365 370375 Gly Glu Asn Gly Lys Met Val Cys Tyr Ile Gln Glu Asn Leu Pro 380 385390 Phe Leu Leu Lys Pro Ser Val Glu Asn Phe Tyr Ile Leu Ile Thr 395 400405 Glu Gly Ala Leu Asp Arg Glu Ile Arg Ala Glu Tyr Asn Ile Thr 410 415420 Ile Thr Val Thr Asp Leu Gly Thr Pro Arg Leu Lys Thr Glu His 425 430435 Asn Ile Thr Val Leu Val Ser Asp Val Asn Asp Asn Ala Pro Ala 440 445450 Phe Thr Gln Thr Ser Tyr Thr Leu Phe Val Arg Glu Asn Asn Ser 455 460465 Pro Ala Leu His Ile Gly Ser Val Ser Ala Thr Asp Arg Asp Ser 470 475480 Gly Thr Asn Ala Gln Val Thr Tyr Ser Leu Leu Pro Pro Gln Asp 485 490495 Pro His Leu Pro Leu Ala Ser Leu Val Ser Ile Asn Ala Asp Asn 500 505510 Gly His Leu Phe Ala Leu Arg Ser Leu Asp Tyr Glu Ala Leu Gln 515 520525 Ala Phe Glu Phe Arg Val Gly Ala Thr Asp Arg Gly Ser Pro Ala 530 535540 Leu Ser Arg Glu Ala Leu Val Arg Val Leu Val Leu Asp Ala Asn 545 550555 Asp Asn Ser Pro Phe Val Leu Tyr Pro Leu Gln Asn Gly Ser Ala 560 565570 Pro Cys Thr Glu Leu Val Pro Arg Ala Ala Glu Pro Gly Tyr Leu 575 580585 Val Thr Lys Val Val Ala Val Asp Gly Asp Ser Gly Gln Asn Ala 590 595600 Trp Leu Ser Tyr Gln Leu Leu Lys Ala Thr Glu Pro Gly Leu Phe 605 610615 Gly Val Trp Ala His Asn Gly Glu Val Arg Thr Ala Arg Leu Leu 620 625630 Ser Glu Arg Asp Ala Ala Lys His Arg Leu Val Val Leu Val Lys 635 640645 Asp Asn Gly Glu Pro Pro Arg Ser Ala Thr Ala Thr Leu His Leu 650 655660 Leu Leu Val Asp Gly Phe Ser Gln Pro Tyr Leu Pro Leu Pro Glu 665 670675 Ala Ala Pro Ala Gln Ala Gln Ala Glu Ala Asp Leu Leu Thr Val 680 685690 Tyr Leu Val Val Ala Leu Ala Ser Val Ser Ser Leu Phe Leu Leu 695 700705 Ser Val Leu Leu Phe Val Ala Val Arg Leu Cys Arg Arg Ser Arg 710 715720 Ala Ala Ser Val Gly Arg Cys Ser Val Pro Glu Gly Pro Phe Pro 725 730735 Gly His Leu Val Asp Val Arg Gly Ala Glu Thr Leu Ser Gln Ser 740 745750 Tyr Gln Tyr Glu Val Cys Leu Thr Gly Gly Pro Gly Thr Ser Glu 755 760765 Phe Lys Phe Leu Lys Pro Val Ile Ser Asp Ile Gln Ala Gln Gly 770 775780 Pro Gly Arg Lys Gly Glu Glu Asn Ser Thr Phe Arg Asn Ser Phe 785 790795 Gly Phe Asn Ile Gln 800 53 24 DNA Artificial Sequence Syntheticoligonucleotide probe 53 ctggggagtg tccttggcag gttc 24 54 27 DNAArtificial Sequence Synthetic oligonucleotide probe 54 cagcatacagggctctttag ggcacac 27 55 46 DNA Artificial Sequence Syntheticoligonucleotide probe 55 cggtgactga ggaaacagag aaaggatcct ttgtggtcaatctggc 46 56 2242 DNA Homo Sapien unsure 2181 unknown base 56 gaatgaatacctccgaagcc gctttgttct ccagatgtga atagctccac 50 tataccagcc tcgtcttccttccgggggac aacgtgggtc agggcacaga 100 gagatattta atgtcaccct cttggggctttcatgggact ccctctgcca 150 cattttttgg aggttgggaa agttgctaga ggcttcagaactccagccta 200 atggatccca aactcgggag aatggctgcg tccctgctgg ctgtgctgct250 gctgctgctg gagcgcggca tgttctcctc accctccccg cccccggcgc 300tgttagagaa agtcttccag tacattgacc tccatcagga tgaatttgtg 350 cagacgctgaaggagtgggt ggccatcgag agcgactctg tccagcctgt 400 gcctcgcttc agacaagagctcttcagaat gatggccgtg gctgcggaca 450 cgctgcagcg cctgggggcc cgtgtggcctcggtggacat gggtcctcag 500 cagctgcccg atggtcagag tcttccaata cctcccgtcatcctggccga 550 actggggagc gatcccacga aaggcaccgt gtgcttctac ggccacttgg600 acgtgcagcc tgctgaccgg ggcgatgggt ggctcacgga cccctatgtg 650ctgacggagg tagacgggaa actttatgga cgaggagcga ccgacaacaa 700 aggccctgtcttggcttgga tcaatgctgt gagcgccttc agagccctgg 750 agcaagatct tcctgtgaatatcaaattca tcattgaggg gatggaagag 800 gctggctctg ttgccctgga ggaacttgtggaaaaagaaa aggaccgatt 850 cttctctggt gtggactaca ttgtaatttc agataacctgtggatcagcc 900 aaaggaagcc agcaatcact tatggaaccc gggggaacag ctacttcatg950 gtggaggtga aatgcagaga ccaggatttt cactcaggaa cctttggtgg 1000catccttcat gaaccaatgg ctgatctggt tgctcttctc ggtagcctgg 1050 tagactcgtctggtcatatc ctggtccctg gaatctatga tgaagtggtt 1100 cctcttacag aagaggaaataaatacatac aaagccatcc atctagacct 1150 agaagaatac cggaatagca gccgggttgagaaatttctg ttcgatacta 1200 aggaggagat tctaatgcac ctctggaggt acccatctctttctattcat 1250 gggatcgagg gcgcgtttga tgagcctgga actaaaacag tcatacctgg1300 ccgagttata ggaaaatttt caatccgtct agtccctcac atgaatgtgt 1350ctgcggtgga aaaacaggtg acacgacatc ttgaagatgt gttctccaaa 1400 agaaatagttccaacaagat ggttgtttcc atgactctag gactacaccc 1450 gtggattgca aatattgatgacacccagta tctcgcagca aaaagagcga 1500 tcagaacagt gtttggaaca gaaccagatatgatccggga tggatccacc 1550 attccaattg ccaaaatgtt ccaggagatc gtccacaagagcgtggtgct 1600 aattccgctg ggagctgttg atgatggaga acattcgcag aatgagaaaa1650 tcaacaggtg gaactacata gagggaacca aattatttgc tgcctttttc 1700ttagagatgg cccagctcca ttaatcacaa gaaccttcta gtctgatctg 1750 atccactgacagattcacct cccccacatc cctagacagg gatggaatgt 1800 aaatatccag agaatttgggtctagtatag tacattttcc cttccattta 1850 aaatgtcttg ggatatctgg atcagtaataaaatatttca aaggcacaga 1900 tgttggaaat ggtttaaggt cccccactgc acaccttcctcaagtcatag 1950 ctgcttgcag caacttgatt tccccaagtc ctgtgcaata gccccaggat2000 tggattcctt ccaacctttt agcatatctc caaccttgca atttgattgg 2050cataatcact ccggtttgct ttctaggtcc tcaagtgctc gtgacacata 2100 atcattccatccaatgatcg cctttgcttt accactcttt ccttttatct 2150 tattaataaa aatgttggtctccaccactg nctcccaaaa aaaaaaaaaa 2200 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaaaaaaaaa aa 2242 57 507 PRT Homo Sapien 57 Met Asp Pro Lys Leu Gly ArgMet Ala Ala Ser Leu Leu Ala Val 1 5 10 15 Leu Leu Leu Leu Leu Glu ArgGly Met Phe Ser Ser Pro Ser Pro 20 25 30 Pro Pro Ala Leu Leu Glu Lys ValPhe Gln Tyr Ile Asp Leu His 35 40 45 Gln Asp Glu Phe Val Gln Thr Leu LysGlu Trp Val Ala Ile Glu 50 55 60 Ser Asp Ser Val Gln Pro Val Pro Arg PheArg Gln Glu Leu Phe 65 70 75 Arg Met Met Ala Val Ala Ala Asp Thr Leu GlnArg Leu Gly Ala 80 85 90 Arg Val Ala Ser Val Asp Met Gly Pro Gln Gln LeuPro Asp Gly 95 100 105 Gln Ser Leu Pro Ile Pro Pro Val Ile Leu Ala GluLeu Gly Ser 110 115 120 Asp Pro Thr Lys Gly Thr Val Cys Phe Tyr Gly HisLeu Asp Val 125 130 135 Gln Pro Ala Asp Arg Gly Asp Gly Trp Leu Thr AspPro Tyr Val 140 145 150 Leu Thr Glu Val Asp Gly Lys Leu Tyr Gly Arg GlyAla Thr Asp 155 160 165 Asn Lys Gly Pro Val Leu Ala Trp Ile Asn Ala ValSer Ala Phe 170 175 180 Arg Ala Leu Glu Gln Asp Leu Pro Val Asn Ile LysPhe Ile Ile 185 190 195 Glu Gly Met Glu Glu Ala Gly Ser Val Ala Leu GluGlu Leu Val 200 205 210 Glu Lys Glu Lys Asp Arg Phe Phe Ser Gly Val AspTyr Ile Val 215 220 225 Ile Ser Asp Asn Leu Trp Ile Ser Gln Arg Lys ProAla Ile Thr 230 235 240 Tyr Gly Thr Arg Gly Asn Ser Tyr Phe Met Val GluVal Lys Cys 245 250 255 Arg Asp Gln Asp Phe His Ser Gly Thr Phe Gly GlyIle Leu His 260 265 270 Glu Pro Met Ala Asp Leu Val Ala Leu Leu Gly SerLeu Val Asp 275 280 285 Ser Ser Gly His Ile Leu Val Pro Gly Ile Tyr AspGlu Val Val 290 295 300 Pro Leu Thr Glu Glu Glu Ile Asn Thr Tyr Lys AlaIle His Leu 305 310 315 Asp Leu Glu Glu Tyr Arg Asn Ser Ser Arg Val GluLys Phe Leu 320 325 330 Phe Asp Thr Lys Glu Glu Ile Leu Met His Leu TrpArg Tyr Pro 335 340 345 Ser Leu Ser Ile His Gly Ile Glu Gly Ala Phe AspGlu Pro Gly 350 355 360 Thr Lys Thr Val Ile Pro Gly Arg Val Ile Gly LysPhe Ser Ile 365 370 375 Arg Leu Val Pro His Met Asn Val Ser Ala Val GluLys Gln Val 380 385 390 Thr Arg His Leu Glu Asp Val Phe Ser Lys Arg AsnSer Ser Asn 395 400 405 Lys Met Val Val Ser Met Thr Leu Gly Leu His ProTrp Ile Ala 410 415 420 Asn Ile Asp Asp Thr Gln Tyr Leu Ala Ala Lys ArgAla Ile Arg 425 430 435 Thr Val Phe Gly Thr Glu Pro Asp Met Ile Arg AspGly Ser Thr 440 445 450 Ile Pro Ile Ala Lys Met Phe Gln Glu Ile Val HisLys Ser Val 455 460 465 Val Leu Ile Pro Leu Gly Ala Val Asp Asp Gly GluHis Ser Gln 470 475 480 Asn Glu Lys Ile Asn Arg Trp Asn Tyr Ile Glu GlyThr Lys Leu 485 490 495 Phe Ala Ala Phe Phe Leu Glu Met Ala Gln Leu His500 505 58 1470 DNA Homo Sapien 58 ctcggctgga tttaaggttg ccgctagccgcctgggaatt taagggaccc 50 acactacctt cccgaagttg aaggcaagcg gtgattgtttgtagacggcg 100 ctttgtcatg ggacctgtgc ggttgggaat attgcttttc ctttttttgg150 ccgtgcacga ggcttgggct gggatgttga aggaggagga cgatgacaca 200gaacgcttgc ccagcaaatg cgaagtgtgt aagctgctga gcacagagct 250 acaggcggaactgagtcgca ccggtcgatc tcgagaggtg ctggagctgg 300 ggcaggtgct ggatacaggcaagaggaaga gacacgtgcc ttacagcgtt 350 tcagagacaa ggctggaaga ggccttagagaatttatgtg agcggatcct 400 ggactatagt gttcacgctg agcgcaaggg ctcactgagatatgccaagg 450 gtcagagtca gaccatggca acactgaaag gcctagtgca gaagggggtg500 aaggtggatc tggggatccc tctggagctt tgggatgagc ccagcgtgga 550ggtcacatac ctcaagaagc agtgtgagac catgttggag gagtttgaag 600 acattgtgggagactggtac ttccaccatc aggagcagcc cctacaaaat 650 tttctctgtg aaggtcatgtgctcccagct gctgaaactg catgtctaca 700 ggaaacttgg actggaaagg agatcacagatggggaagag aaaacagaag 750 gggaggaaga gcaggaggag gaggaggaag aggaggaagaggaaggggga 800 gacaagatga ccaagacagg aagccacccc aaacttgacc gagaagatct850 ttgacccttg cctttgagcc cccaggaggg gaagggatca tggagagccc 900tctaaagcct gcactctccc tgctccacag ctttcagggt gtgtttatga 950 gtgactccacccaagcttgt agctgttctc tcccatctaa cctcaggcaa 1000 gatcctggtg aaacagcatgacatggcttc tggggtggag ggtgggggtg 1050 gaggtcctgc tcctagagat gaactctatccagcccctta attggcaggt 1100 gtatgtgctg acagtactga aagctttcct ctttaactgatcccaccccc 1150 acccaaaagt cagcagtggc actggagctg tgggctttgg ggaagtcact1200 tagctcctta aggtctgttt ttagaccctt ccaaggaaga ggccagaacg 1250gacattctct gcgatctata tacattgcct gtatccagga ggctacacac 1300 cagcaaaccgtgaaggagaa tgggacactg ggtcatggcc tggagttgct 1350 gataatttag gtgggatagatacttggtct acttaagctc aatgtaaccc 1400 agagcccacc atatagtttt ataggtgctcaactttctat atcgctatta 1450 aacttttttc tttttttcta 1470 59 248 PRT HomoSapien 59 Met Gly Pro Val Arg Leu Gly Ile Leu Leu Phe Leu Phe Leu Ala 15 10 15 Val His Glu Ala Trp Ala Gly Met Leu Lys Glu Glu Asp Asp Asp 2025 30 Thr Glu Arg Leu Pro Ser Lys Cys Glu Val Cys Lys Leu Leu Ser 35 4045 Thr Glu Leu Gln Ala Glu Leu Ser Arg Thr Gly Arg Ser Arg Glu 50 55 60Val Leu Glu Leu Gly Gln Val Leu Asp Thr Gly Lys Arg Lys Arg 65 70 75 HisVal Pro Tyr Ser Val Ser Glu Thr Arg Leu Glu Glu Ala Leu 80 85 90 Glu AsnLeu Cys Glu Arg Ile Leu Asp Tyr Ser Val His Ala Glu 95 100 105 Arg LysGly Ser Leu Arg Tyr Ala Lys Gly Gln Ser Gln Thr Met 110 115 120 Ala ThrLeu Lys Gly Leu Val Gln Lys Gly Val Lys Val Asp Leu 125 130 135 Gly IlePro Leu Glu Leu Trp Asp Glu Pro Ser Val Glu Val Thr 140 145 150 Tyr LeuLys Lys Gln Cys Glu Thr Met Leu Glu Glu Phe Glu Asp 155 160 165 Ile ValGly Asp Trp Tyr Phe His His Gln Glu Gln Pro Leu Gln 170 175 180 Asn PheLeu Cys Glu Gly His Val Leu Pro Ala Ala Glu Thr Ala 185 190 195 Cys LeuGln Glu Thr Trp Thr Gly Lys Glu Ile Thr Asp Gly Glu 200 205 210 Glu LysThr Glu Gly Glu Glu Glu Gln Glu Glu Glu Glu Glu Glu 215 220 225 Glu GluGlu Glu Gly Gly Asp Lys Met Thr Lys Thr Gly Ser His 230 235 240 Pro LysLeu Asp Arg Glu Asp Leu 245 60 890 DNA Homo Sapien 60 aagtacttgtgtccgggtgg tggactggat tagctgcgga gccctggaag 50 ctgcctgtcc ttctccctgtgcttaaccag aggtgcccat gggttggaca 100 atgaggctgg tcacagcagc actgttactgggtctcatga tggtggtcac 150 tggagacgag gatgagaaca gcccgtgtgc ccatgaggccctcttggacg 200 aggacaccct cttttgccag ggccttgaag ttttctaccc agagttgggg250 aacattggct gcaaggttgt tcctgattgt aacaactaca gacagaagat 300cacctcctgg atggagccga tagtcaagtt cccgggggcc gtggacggcg 350 caacctatatcctggtgatg gtggatccag atgcccctag cagagcagaa 400 cccagacaga gattctggagacattggctg gtaacagata tcaagggcgc 450 cgacctgaag aaagggaaga ttcagggccaggagttatca gcctaccagg 500 ctccctcccc accggcacac agtggcttcc atcgctaccagttctttgtc 550 tatcttcagg aaggaaaagt catctctctc cttcccaagg aaaacaaaac600 tcgaggctct tggaaaatgg acagatttct gaaccgcttc cacctgggcg 650aacctgaagc aagcacccag ttcatgaccc agaactacca ggactcacca 700 accctccaggctcccagagg aagggccagc gagcccaagc acaaaaccag 750 gcagagatag ctgcctgctagatagccggc tttgccatcc gggcatgtgg 800 ccacactgct caccaccgac gatgtgggtatggaaccccc tctggataca 850 aaccccttc ttttccaaat taaaaaaaaa aatcatcaaa 89061 223 PRT Homo Sapien 61 Met Gly Trp Thr Met Arg Leu Val Thr Ala AlaLeu Leu Leu Gly 1 5 10 15 Leu Met Met Val Val Thr Gly Asp Glu Asp GluAsn Ser Pro Cys 20 25 30 Ala His Glu Ala Leu Leu Asp Glu Asp Thr Leu PheCys Gln Gly 35 40 45 Leu Glu Val Phe Tyr Pro Glu Leu Gly Asn Ile Gly CysLys Val 50 55 60 Val Pro Asp Cys Asn Asn Tyr Arg Gln Lys Ile Thr Ser TrpMet 65 70 75 Glu Pro Ile Val Lys Phe Pro Gly Ala Val Asp Gly Ala Thr Tyr80 85 90 Ile Leu Val Met Val Asp Pro Asp Ala Pro Ser Arg Ala Glu Pro 95100 105 Arg Gln Arg Phe Trp Arg His Trp Leu Val Thr Asp Ile Lys Gly 110115 120 Ala Asp Leu Lys Lys Gly Lys Ile Gln Gly Gln Glu Leu Ser Ala 125130 135 Tyr Gln Ala Pro Ser Pro Pro Ala His Ser Gly Phe His Arg Tyr 140145 150 Gln Phe Phe Val Tyr Leu Gln Glu Gly Lys Val Ile Ser Leu Leu 155160 165 Pro Lys Glu Asn Lys Thr Arg Gly Ser Trp Lys Met Asp Arg Phe 170175 180 Leu Asn Arg Phe His Leu Gly Glu Pro Glu Ala Ser Thr Gln Phe 185190 195 Met Thr Gln Asn Tyr Gln Asp Ser Pro Thr Leu Gln Ala Pro Arg 200205 210 Gly Arg Ala Ser Glu Pro Lys His Lys Thr Arg Gln Arg 215 220 621321 DNA Homo Sapien 62 gtcgacccac gcgtccgaag ctgctggagc cacgattcagtcccctggac 50 tgtagataaa gaccctttct tgccaggtgc tgagacaacc acactatgag 100aggcactcca ggagacgctg atggtggagg aagggccgtc tatcaatcaa 150 tcactgttgctgttatcaca tgcaagtatc cagaggctct tgagcaaggc 200 agaggggatc ccatttatttgggaatccag aatccagaaa tgtgtttgta 250 ttgtgagaag gttggagaac agcccacattgcagctaaaa gagcagaaga 300 tcatggatct gtatggccaa cccgagcccg tgaaacccttccttttctac 350 cgtgccaaga ctggtaggac ctccaccctt gagtctgtgg ccttcccgga400 ctggttcatt gcctcctcca agagagacca gcccatcatt ctgacttcag 450aacttgggaa gtcatacaac actgcctttg aattaaatat aaatgactga 500 actcagcctagaggtggcag cttggtcttt gtcttaaagt ttctggttcc 550 caatgtgttt tcgtctacattttcttagtg tcattttcac gctggtgctg 600 agacaggagc aaggctgctg ttatcatctcattttataat gaagaagaag 650 caattacttc atagcaactg aagaacagga tgtggcctcagaagcaggag 700 agctgggtgg tataaggctg tcctctcaag ctggtgctgt gtaggccaca750 aggcatctgc atgagtgact ttaagactca aagaccaaac actgagcttt 800cttctagggg tgggtatgaa gatgcttcag agctcatgcg cgttacccac 850 gatggcatgactagcacaga gctgatctct gtttctgttt tgctttattc 900 cctcttggga tgatatcatccagtctttat atgttgccaa tatacctcat 950 tgtgtgtaat agaaccttct tagcattaagaccttgtaaa caaaaataat 1000 tcttggggtg ggtatgaaga tgcttcagag ctcatgcgcgttacccacga 1050 tggcatgact agcacagagc tgatctctgt ttctgttttg ctttattccc1100 tcttgggatg atatcatcca gtctttatat gttgccaata tacctcattg 1150tgtgtaatag aaccttctta gcattaagac cttgtaaaca aaaataattc 1200 ttgtgttaagttaaatcatt tttgtcctaa ttgtaatgtg taatcttaaa 1250 gttaaataaa ctttgtgtatttatataata ataaagctaa aactgatata 1300 aaataaagaa agagtaaact g 1321 63134 PRT Homo Sapien 63 Met Arg Gly Thr Pro Gly Asp Ala Asp Gly Gly GlyArg Ala Val 1 5 10 15 Tyr Gln Ser Ile Thr Val Ala Val Ile Thr Cys LysTyr Pro Glu 20 25 30 Ala Leu Glu Gln Gly Arg Gly Asp Pro Ile Tyr Leu GlyIle Gln 35 40 45 Asn Pro Glu Met Cys Leu Tyr Cys Glu Lys Val Gly Glu GlnPro 50 55 60 Thr Leu Gln Leu Lys Glu Gln Lys Ile Met Asp Leu Tyr Gly Gln65 70 75 Pro Glu Pro Val Lys Pro Phe Leu Phe Tyr Arg Ala Lys Thr Gly 8085 90 Arg Thr Ser Thr Leu Glu Ser Val Ala Phe Pro Asp Trp Phe Ile 95 100105 Ala Ser Ser Lys Arg Asp Gln Pro Ile Ile Leu Thr Ser Glu Leu 110 115120 Gly Lys Ser Tyr Asn Thr Ala Phe Glu Leu Asn Ile Asn Asp 125 130 64999 DNA Homo Sapien 64 gcgaggctgc accagcgcct ggcaccatga ggacgcctgggcctctgccc 50 gtgctgctgc tgctcctggc gggagccccc gccgcgcggc ccactccccc 100gacctgctac tcccgcatgc gggccctgag ccaggagatc acccgcgact 150 tcaacctcctgcaggtctcg gagccctcgg agccatgtgt gagatacctg 200 cccaggctgt acctggacatacacaattac tgtgtgctgg acaagctgcg 250 ggactttgtg gcctcgcccc cgtgttggaaagtggcccag gtagattcct 300 tgaaggacaa agcacggaag ctgtacacca tcatgaactcgttctgcagg 350 agagatttgg tattcctgtt ggatgactgc aatgccttgg aatacccaat400 cccagtgact acggtcctgc cagatcgtca gcgctaaggg aactgagacc 450agagaaagaa cccaagagaa ctaaagttat gtcagctacc cagacttaat 500 gggccagagccatgaccctc acaggtcttg tgttagttgt atctgaaact 550 gttatgtatc tctctaccttctggaaaaca gggctggtat tcctacccag 600 gaacctcctt tgagcataga gttagcaaccatgcttctca ttcccttgac 650 tcatgtcttg ccaggatggt tagatacaca gcatgttgatttggtcacta 700 aaaagaagaa aaggactaac aagcttcact tttatgaaca actattttga750 gaacatgcac aatagtatgt ttttattact ggtttaatgg agtaatggta 800cttttattct ttcttgatag aaacctgctt acatttaacc aagcttctat 850 tatgcctttttctaacacag actttcttca ctgtctttca tttaaaaaga 900 aattaatgct cttaagatatatattttacg tagtgctgac aggacccact 950 ctttcattga aaggtgatga aaatcaaataaagaatctct tcacatgga 999 65 136 PRT Homo Sapien 65 Met Arg Thr Pro GlyPro Leu Pro Val Leu Leu Leu Leu Leu Ala 1 5 10 15 Gly Ala Pro Ala AlaArg Pro Thr Pro Pro Thr Cys Tyr Ser Arg 20 25 30 Met Arg Ala Leu Ser GlnGlu Ile Thr Arg Asp Phe Asn Leu Leu 35 40 45 Gln Val Ser Glu Pro Ser GluPro Cys Val Arg Tyr Leu Pro Arg 50 55 60 Leu Tyr Leu Asp Ile His Asn TyrCys Val Leu Asp Lys Leu Arg 65 70 75 Asp Phe Val Ala Ser Pro Pro Cys TrpLys Val Ala Gln Val Asp 80 85 90 Ser Leu Lys Asp Lys Ala Arg Lys Leu TyrThr Ile Met Asn Ser 95 100 105 Phe Cys Arg Arg Asp Leu Val Phe Leu LeuAsp Asp Cys Asn Ala 110 115 120 Leu Glu Tyr Pro Ile Pro Val Thr Thr ValLeu Pro Asp Arg Gln 125 130 135 Arg 66 1893 DNA Homo Sapien 66gtctccgcgt cacaggaact tcagcaccca cagggcggac agcgctcccc 50 tctacctggagacttgactc ccgcgcgccc caaccctgct tatcccttga 100 ccgtcgagtg tcagagatcctgcagccgcc cagtcccggc ccctctcccg 150 ccccacaccc accctcctgg ctcttcctgtttttactcct ccttttcatt 200 cataacaaaa gctacagctc caggagccca gcgccgggctgtgacccaag 250 ccgagcgtgg aagaatgggg ttcctcggga ccggcacttg gattctggtg300 ttagtgctcc cgattcaagc tttccccaaa cctggaggaa gccaagacaa 350atctctacat aatagagaat taagtgcaga aagacctttg aatgaacaga 400 ttgctgaagcagaagaagac aagattaaaa aaacatatcc tccagaaaac 450 aagccaggtc agagcaactattcttttgtt gataacttga acctgctaaa 500 ggcaataaca gaaaaggaaa aaattgagaaagaaagacaa tctataagaa 550 gctccccact tgataataag ttgaatgtgg aagatgttgattcaaccaag 600 aatcgaaaac tgatcgatga ttatgactct actaagagtg gattggatca650 taaatttcaa gatgatccag atggtcttca tcaactagac gggactcctt 700taaccgctga agacattgtc cataaaatcg ctgccaggat ttatgaagaa 750 aatgacagagccgtgtttga caagattgtt tctaaactac ttaatctcgg 800 ccttatcaca gaaagccaagcacatacact ggaagatgaa gtagcagagg 850 ttttacaaaa attaatctca aaggaagccaacaattatga ggaggatccc 900 aataagccca caagctggac tgagaatcag gctggaaaaataccagagaa 950 agtgactcca atggcagcaa ttcaagatgg tcttgctaag ggagaaaacg1000 atgaaacagt atctaacaca ttaaccttga caaatggctt ggaaaggaga 1050actaaaacct acagtgaaga caactttgag gaactccaat atttcccaaa 1100 tttctatgcgctactgaaaa gtattgattc agaaaaagaa gcaaaagaga 1150 aagaaacact gattactatcatgaaaacac tgattgactt tgtgaagatg 1200 atggtgaaat atggaacaat atctccagaagaaggtgttt cctaccttga 1250 aaacttggat gaaatgattg ctcttcagac caaaaacaagctagaaaaaa 1300 atgctactga caatataagc aagcttttcc cagcaccatc agagaagagt1350 catgaagaaa cagacagtac caaggaagaa gcagctaaga tggaaaagga 1400atatggaagc ttgaaggatt ccacaaaaga tgataactcc aacccaggag 1450 gaaagacagatgaacccaaa ggaaaaacag aagcctattt ggaagccatc 1500 agaaaaaata ttgaatggttgaagaaacat gacaaaaagg gaaataaaga 1550 agattatgac ctttcaaaga tgagagacttcatcaataaa caagctgatg 1600 cttatgtgga gaaaggcatc cttgacaagg aagaagccgaggccatcaag 1650 cgcatttata gcagcctgta aaaatggcaa aagatccagg agtctttcaa1700 ctgtttcaga aaacataata tagcttaaaa cacttctaat tctgtgatta 1750aaattttttg acccaagggt tattagaaag tgctgaattt acagtagtta 1800 accttttacaagtggttaaa acatagcttt cttcccgtaa aaactatctg 1850 aagtaaagt tgtatgtaagctgaaaaaaa aaaaaaaaaa aaa 1893 67 468 PRT Homo Sapien 67 Met Gly Phe LeuGly Thr Gly Thr Trp Ile Leu Val Leu Val Leu 1 5 10 15 Pro Ile Gln AlaPhe Pro Lys Pro Gly Gly Ser Gln Asp Lys Ser 20 25 30 Leu His Asn Arg GluLeu Ser Ala Glu Arg Pro Leu Asn Glu Gln 35 40 45 Ile Ala Glu Ala Glu GluAsp Lys Ile Lys Lys Thr Tyr Pro Pro 50 55 60 Glu Asn Lys Pro Gly Gln SerAsn Tyr Ser Phe Val Asp Asn Leu 65 70 75 Asn Leu Leu Lys Ala Ile Thr GluLys Glu Lys Ile Glu Lys Glu 80 85 90 Arg Gln Ser Ile Arg Ser Ser Pro LeuAsp Asn Lys Leu Asn Val 95 100 105 Glu Asp Val Asp Ser Thr Lys Asn ArgLys Leu Ile Asp Asp Tyr 110 115 120 Asp Ser Thr Lys Ser Gly Leu Asp HisLys Phe Gln Asp Asp Pro 125 130 135 Asp Gly Leu His Gln Leu Asp Gly ThrPro Leu Thr Ala Glu Asp 140 145 150 Ile Val His Lys Ile Ala Ala Arg IleTyr Glu Glu Asn Asp Arg 155 160 165 Ala Val Phe Asp Lys Ile Val Ser LysLeu Leu Asn Leu Gly Leu 170 175 180 Ile Thr Glu Ser Gln Ala His Thr LeuGlu Asp Glu Val Ala Glu 185 190 195 Val Leu Gln Lys Leu Ile Ser Lys GluAla Asn Asn Tyr Glu Glu 200 205 210 Asp Pro Asn Lys Pro Thr Ser Trp ThrGlu Asn Gln Ala Gly Lys 215 220 225 Ile Pro Glu Lys Val Thr Pro Met AlaAla Ile Gln Asp Gly Leu 230 235 240 Ala Lys Gly Glu Asn Asp Glu Thr ValSer Asn Thr Leu Thr Leu 245 250 255 Thr Asn Gly Leu Glu Arg Arg Thr LysThr Tyr Ser Glu Asp Asn 260 265 270 Phe Glu Glu Leu Gln Tyr Phe Pro AsnPhe Tyr Ala Leu Leu Lys 275 280 285 Ser Ile Asp Ser Glu Lys Glu Ala LysGlu Lys Glu Thr Leu Ile 290 295 300 Thr Ile Met Lys Thr Leu Ile Asp PheVal Lys Met Met Val Lys 305 310 315 Tyr Gly Thr Ile Ser Pro Glu Glu GlyVal Ser Tyr Leu Glu Asn 320 325 330 Leu Asp Glu Met Ile Ala Leu Gln ThrLys Asn Lys Leu Glu Lys 335 340 345 Asn Ala Thr Asp Asn Ile Ser Lys LeuPhe Pro Ala Pro Ser Glu 350 355 360 Lys Ser His Glu Glu Thr Asp Ser ThrLys Glu Glu Ala Ala Lys 365 370 375 Met Glu Lys Glu Tyr Gly Ser Leu LysAsp Ser Thr Lys Asp Asp 380 385 390 Asn Ser Asn Pro Gly Gly Lys Thr AspGlu Pro Lys Gly Lys Thr 395 400 405 Glu Ala Tyr Leu Glu Ala Ile Arg LysAsn Ile Glu Trp Leu Lys 410 415 420 Lys His Asp Lys Lys Gly Asn Lys GluAsp Tyr Asp Leu Ser Lys 425 430 435 Met Arg Asp Phe Ile Asn Lys Gln AlaAsp Ala Tyr Val Glu Lys 440 445 450 Gly Ile Leu Asp Lys Glu Glu Ala GluAla Ile Lys Arg Ile Tyr 455 460 465 Ser Ser Leu 68 22 DNA ArtificialSequence Synthetic oligonucleotide probe 68 cgtcacagga acttcagcac cc 2269 23 DNA Artificial Sequence Synthetic oligonucleotide probe 69gtcttggctt cctccaggtt tgg 23 70 38 DNA Artificial Sequence Syntheticoligonucleotide probe 70 ggacagcgct cccctctacc tggagacttg actcccgc 38 712379 DNA Homo Sapien 71 gttgctccgg cggcgctcgg ggagggagcc agcagcctagggcctaggcc 50 cgggccacca tggcgctgcc tccaggccca gccgccctcc ggcacacact 100gctgctcctg ccagcccttc tgagctcagg ttggggggag ttggagccac 150 aaatagatggtcagacctgg gctgagcggg cacttcggga gaatgaacgc 200 cacgccttca cctgccgggtggcagggggg cctggcaccc ccagattggc 250 ctggtatctg gatggacagc tgcaggaggccagcacctca agactgctga 300 gcgtgggagg ggaggccttc tctggaggca ccagcaccttcactgtcact 350 gcccatcggg cccagcatga gctcaactgc tctctgcagg accccagaag400 tggccgatca gccaacgcct ctgtcatcct taatgtgcaa ttcaagccag 450agattgccca agtcggcgcc aagtaccagg aagctcaggg cccaggcctc 500 ctggttgtcctgtttgccct ggtgcgtgcc aacccgccgg ccaatgtcac 550 ctggatcgac caggatgggccagtgactgt caacacctct gacttcctgg 600 tgctggatgc gcagaactac ccctggctcaccaaccacac ggtgcagctg 650 cagctccgca gcctggcaca caacctctcg gtggtggccaccaatgacgt 700 gggtgtcacc agtgcgtcgc ttccagcccc aggcccctcc cggcacccat750 ctctgatatc aagtgactcc aacaacctaa aactcaacaa cgtgcgcctg 800ccacgggaga acatgtccct cccgtccaac cttcagctca atgacctcac 850 tccagattccagagcagtga aaccagcaga ccggcagatg gctcagaaca 900 acagccggcc agagcttctggacccggagc ccggcggcct cctcaccagc 950 caaggtttca tccgcctccc agtgctgggctatatctatc gagtgtccag 1000 cgtgagcagt gatgagatct ggctctgagc cgagggcgagacaggagtat 1050 tctcttggcc tctggacacc ctcccattcc tccaaggcat cctctaccta1100 gctaggtcac caacgtgaag aagttatgcc actgccactt ttgcttgccc 1150tcctggctgg ggtgccctcc atgtcatgca cgtgatgcat ttcactgggc 1200 tgtaacccgcaggggcacag gtatctttgg caaggctacc agttggacgt 1250 aagcccctca tgctgactcagggtgggccc tgcatgtgat gactgggccc 1300 ttccagaggg agctctttgg ccaggggtgttcagatgtca tccagcatcc 1350 aagtgtggca tggcctgctg tataccccac cccagtactccacagcacct 1400 tgtacagtag gcatgggggc gtgcctgtgt gggggacagg gagggccctg1450 catggatttt cctccttcct atgctatgta gccttgttcc ctcaggtaaa 1500atttaggacc ctgctagctg tgcagaaccc aattgccctt tgcacagaaa 1550 ccaacccctgacccagcggt accggccaag cacaaacgtc ctttttgctg 1600 cacacgtctc tgcccttcacttcttctctt ctgtccccac ctcctcttgg 1650 gaattctagg ttacacgttg gaccttctctactacttcac tgggcactag 1700 acttttctat tggcctgtgc catcgcccag tattagcacaagttagggag 1750 gaagaggcag gcgatgagtc tagtagcacc caggacggct tgtagctatg1800 catcattttc ctacggcgtt agcactttaa gcacatcccc taggggaggg 1850ggtgagtgag gggcccagag ccctctttgt ggcttcccca cgtttggcct 1900 tctgggattcactgtgagtg tcctgagctc tcggggttga tggtttttct 1950 ctcagcatgt ctcctccaccacgggacccc agccctgacc aacccatggt 2000 tgcctcatca gcaggaaggt gcccttcctggaggatggtc gccacaggca 2050 cataattcaa cagtgtggaa gctttagggg aacatggagaaagaaggaga 2100 ccacataccc caaagtgacc taagaacact ttaaaaagca acatgtaaat2150 gattggaaat taatatagta cagaatatat ttttcccttg ttgagatctt 2200cttttgtaat gtttttcatg ttactgccta gggcggtgct gagcacacag 2250 caagtttaataaacttgact gaattcattt aaaaaaaaaa aaaaaaaaaa 2300 aaaaaaaaaa aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2350 aaaaaaaaaa aaaaaaaaaa aaaaaaaaa2379 72 322 PRT Homo Sapien 72 Met Ala Leu Pro Pro Gly Pro Ala Ala LeuArg His Thr Leu Leu 1 5 10 15 Leu Leu Pro Ala Leu Leu Ser Ser Gly TrpGly Glu Leu Glu Pro 20 25 30 Gln Ile Asp Gly Gln Thr Trp Ala Glu Arg AlaLeu Arg Glu Asn 35 40 45 Glu Arg His Ala Phe Thr Cys Arg Val Ala Gly GlyPro Gly Thr 50 55 60 Pro Arg Leu Ala Trp Tyr Leu Asp Gly Gln Leu Gln GluAla Ser 65 70 75 Thr Ser Arg Leu Leu Ser Val Gly Gly Glu Ala Phe Ser GlyGly 80 85 90 Thr Ser Thr Phe Thr Val Thr Ala His Arg Ala Gln His Glu Leu95 100 105 Asn Cys Ser Leu Gln Asp Pro Arg Ser Gly Arg Ser Ala Asn Ala110 115 120 Ser Val Ile Leu Asn Val Gln Phe Lys Pro Glu Ile Ala Gln Val125 130 135 Gly Ala Lys Tyr Gln Glu Ala Gln Gly Pro Gly Leu Leu Val Val140 145 150 Leu Phe Ala Leu Val Arg Ala Asn Pro Pro Ala Asn Val Thr Trp155 160 165 Ile Asp Gln Asp Gly Pro Val Thr Val Asn Thr Ser Asp Phe Leu170 175 180 Val Leu Asp Ala Gln Asn Tyr Pro Trp Leu Thr Asn His Thr Val185 190 195 Gln Leu Gln Leu Arg Ser Leu Ala His Asn Leu Ser Val Val Ala200 205 210 Thr Asn Asp Val Gly Val Thr Ser Ala Ser Leu Pro Ala Pro Gly215 220 225 Pro Ser Arg His Pro Ser Leu Ile Ser Ser Asp Ser Asn Asn Leu230 235 240 Lys Leu Asn Asn Val Arg Leu Pro Arg Glu Asn Met Ser Leu Pro245 250 255 Ser Asn Leu Gln Leu Asn Asp Leu Thr Pro Asp Ser Arg Ala Val260 265 270 Lys Pro Ala Asp Arg Gln Met Ala Gln Asn Asn Ser Arg Pro Glu275 280 285 Leu Leu Asp Pro Glu Pro Gly Gly Leu Leu Thr Ser Gln Gly Phe290 295 300 Ile Arg Leu Pro Val Leu Gly Tyr Ile Tyr Arg Val Ser Ser Val305 310 315 Ser Ser Asp Glu Ile Trp Leu 320 73 843 DNA Homo Sapien 73cggggacgga agcggcccct gggcccgagg ggctggagcc gggccggggc 50 gatgtggagcgcgggccgcg gcggggctgc ctggccggtg ctgttggggc 100 tgctgctggc gctgttagtgccgggcggtg gtgccgccaa gaccggtgcg 150 gagctcgtga cctgcgggtc ggtgctgaagctgctcaata cgcaccaccg 200 cgtgcggctg cactcgcacg acatcaaata cggatccggcagcggccagc 250 aatcggtgac cggcgtagag gcgtcggacg acgccaatag ctactggcgg300 atccgcggcg gctcggaggg cgggtgcccg cgcgggtccc cggtgcgctg 350cgggcaggcg gtgaggctca cgcatgtgct tacgggcaag aacctgcaca 400 cgcaccacttcccgtcgccg ctgtccaaca accaggaggt gagtgccttt 450 ggggaagacg gcgagggcgacgacctggac ctatggacag tgcgctgctc 500 tggacagcac tgggagcgtg aggctgctgtgcgcttccag catgtgggca 550 cctctgtgtt cctgtcagtc acgggtgagc agtatggaagccccatccgt 600 gggcagcatg aggtccacgg catgcccagt gccaacacgc acaatacgtg650 gaaggccatg gaaggcatct tcatcaagcc tagtgtggag ccctctgcag 700gtcacgatga actctgagtg tgtggatgga tgggtggatg gagggtggca 750 ggtggggcgtctgcagggcc actcttggca gagactttgg gtttgtaggg 800 gtcctcaagt gcctttgtgattaaagaatg ttggtctatg aaa 843 74 221 PRT Homo Sapien 74 Met Trp Ser AlaGly Arg Gly Gly Ala Ala Trp Pro Val Leu Leu 1 5 10 15 Gly Leu Leu LeuAla Leu Leu Val Pro Gly Gly Gly Ala Ala Lys 20 25 30 Thr Gly Ala Glu LeuVal Thr Cys Gly Ser Val Leu Lys Leu Leu 35 40 45 Asn Thr His His Arg ValArg Leu His Ser His Asp Ile Lys Tyr 50 55 60 Gly Ser Gly Ser Gly Gln GlnSer Val Thr Gly Val Glu Ala Ser 65 70 75 Asp Asp Ala Asn Ser Tyr Trp ArgIle Arg Gly Gly Ser Glu Gly 80 85 90 Gly Cys Pro Arg Gly Ser Pro Val ArgCys Gly Gln Ala Val Arg 95 100 105 Leu Thr His Val Leu Thr Gly Lys AsnLeu His Thr His His Phe 110 115 120 Pro Ser Pro Leu Ser Asn Asn Gln GluVal Ser Ala Phe Gly Glu 125 130 135 Asp Gly Glu Gly Asp Asp Leu Asp LeuTrp Thr Val Arg Cys Ser 140 145 150 Gly Gln His Trp Glu Arg Glu Ala AlaVal Arg Phe Gln His Val 155 160 165 Gly Thr Ser Val Phe Leu Ser Val ThrGly Glu Gln Tyr Gly Ser 170 175 180 Pro Ile Arg Gly Gln His Glu Val HisGly Met Pro Ser Ala Asn 185 190 195 Thr His Asn Thr Trp Lys Ala Met GluGly Ile Phe Ile Lys Pro 200 205 210 Ser Val Glu Pro Ser Ala Gly His AspGlu Leu 215 220 75 1049 DNA Homo Sapien 75 gttgctatgt tgcccaggctggtcttgaag tgccttgacc tcctaaagtg 50 ttggaaccac agacgtgagc cactccacccagcctaaaac ttcatcttct 100 ttggatgaga tgaacacttt taacaagaga acaggactctatataaatcg 150 ctgtgggctc accacctcta aggaggagca ctgactgaag acagaaaaat200 tgatgaactg aagaagacat ggtccattat gccttacaaa cttacacagt 250gctttgggaa ttccaaagta ctcagtggag agaggtgttt caggagccgt 300 agagccagatcgtcatcatg tctgcattgt ggctgctgct gggcctcctt 350 gccctgatgg acttgtctgaaagcagcaac tggggatgct atggaaacat 400 ccaaagcctg gacacccctg gagcatcttgtgggattgga agacgtcacg 450 gcctgaacta ctgtggagtt cgtgcttctg aaaggctggctgaaatagac 500 atgccatacc tcctgaaata tcaacccatg atgcaaacca ttggccaaaa550 gtactgcatg gatcctgccg tgatcgctgg tgtcttgtcc aggaagtctc 600ccggtgacaa aattctggtc aacatgggcg ataggactag catggtgcag 650 gaccctggctctcaagctcc cacatcctgg attagtgagt ctcaggtttc 700 ccagacaact gaagttctgactactagaat caaagaaatc cagaggaggt 750 ttccaacctg gacccctgac cagtacctgagaggtggact ctgtgcctac 800 agtgggggtg ctggctatgt ccgaagcagc caggacctgagctgtgactt 850 ctgcaatgat gtccttgcac gagccaagta cctcaagaga catggcttct900 aacatctcag atgaaaccca agaccatgat cacatatgca gcctcaaatg 950ttacacagat aaaactagcc aagggcacct gtaactggga atctgagttt 1000 gacctaaaagtcattaaaat aacatgaatc ccattaaaaa aaaaaaaaa 1049 76 194 PRT Homo Sapien76 Met Ser Ala Leu Trp Leu Leu Leu Gly Leu Leu Ala Leu Met Asp 1 5 10 15Leu Ser Glu Ser Ser Asn Trp Gly Cys Tyr Gly Asn Ile Gln Ser 20 25 30 LeuAsp Thr Pro Gly Ala Ser Cys Gly Ile Gly Arg Arg His Gly 35 40 45 Leu AsnTyr Cys Gly Val Arg Ala Ser Glu Arg Leu Ala Glu Ile 50 55 60 Asp Met ProTyr Leu Leu Lys Tyr Gln Pro Met Met Gln Thr Ile 65 70 75 Gly Gln Lys TyrCys Met Asp Pro Ala Val Ile Ala Gly Val Leu 80 85 90 Ser Arg Lys Ser ProGly Asp Lys Ile Leu Val Asn Met Gly Asp 95 100 105 Arg Thr Ser Met ValGln Asp Pro Gly Ser Gln Ala Pro Thr Ser 110 115 120 Trp Ile Ser Glu SerGln Val Ser Gln Thr Thr Glu Val Leu Thr 125 130 135 Thr Arg Ile Lys GluIle Gln Arg Arg Phe Pro Thr Trp Thr Pro 140 145 150 Asp Gln Tyr Leu ArgGly Gly Leu Cys Ala Tyr Ser Gly Gly Ala 155 160 165 Gly Tyr Val Arg SerSer Gln Asp Leu Ser Cys Asp Phe Cys Asn 170 175 180 Asp Val Leu Ala ArgAla Lys Tyr Leu Lys Arg His Gly Phe 185 190 77 899 DNA Homo Sapien 77ttgaaaatct actctatcag ctgctgtggt tgccaccatt ctcaggaccc 50 tcgccatgaaagcccttatg ctgctcaccc tgtctgttct gctctgctgg 100 gtctcagctg acattcgctgtcactcctgc tacaaggtcc ctgtgctggg 150 ctgtgtggac cggcagtcct gccgcctggagccaggacag caatgcctga 200 caacacatgc ataccttggt aagatgtggg ttttctccaatctgcgctgt 250 ggcacaccag aagagccctg tcaggaggcc ttcaaccaaa ccaaccgcaa300 gctgggtctg acatataaca ccacctgctg caacaaggac aactgcaaca 350gcgcaggacc ccggcccact ccagccctgg gccttgtctt ccttacctcc 400 ttggctggccttggcctctg gctgctgcac tgagactcat tccattggct 450 gcccctcctc ccacctgccttggcctgagc ctctctccct gtgtctctgt 500 atcccctggc tttacagaat cgtctctccctagctcccat ttctttaatt 550 aaacactgtt ccgagtggtc tcctcatcca tccttcccacctcacaccct 600 tcactctcct ttttctgggt cccttcccac ttccttccag gacctccatt650 ggctcctaga agggctcccc actttgcttc ctatactctg ctgtccccta 700cttgaggagg gattgggatc tgggcctgaa atggggcttc tgtgttgtcc 750 ccagtgaaggctcccacaag gacctgatga cctcactgta cagagctgac 800 tccccaaacc caggctcccatatgtacccc atcccccata ctcacctctt 850 tccattttga gtaataaatg tctgagtctggaaaaaaaaa aaaaaaaaa 899 78 125 PRT Homo Sapien 78 Met Lys Ala Leu MetLeu Leu Thr Leu Ser Val Leu Leu Cys Trp 1 5 10 15 Val Ser Ala Asp IleArg Cys His Ser Cys Tyr Lys Val Pro Val 20 25 30 Leu Gly Cys Val Asp ArgGln Ser Cys Arg Leu Glu Pro Gly Gln 35 40 45 Gln Cys Leu Thr Thr His AlaTyr Leu Gly Lys Met Trp Val Phe 50 55 60 Ser Asn Leu Arg Cys Gly Thr ProGlu Glu Pro Cys Gln Glu Ala 65 70 75 Phe Asn Gln Thr Asn Arg Lys Leu GlyLeu Thr Tyr Asn Thr Thr 80 85 90 Cys Cys Asn Lys Asp Asn Cys Asn Ser AlaGly Pro Arg Pro Thr 95 100 105 Pro Ala Leu Gly Leu Val Phe Leu Thr SerLeu Ala Gly Leu Gly 110 115 120 Leu Trp Leu Leu His 125 79 1977 DNA HomoSapien 79 acgggccgca gcggcagtga cgtagggttg gcgcacggat ccgttgcggc 50tgcagctctg cagtcgggcc gttccttcgc cgccgccagg ggtagcggtg 100 tagctgcgcagcgtcgcgcg cgctaccgca cccaggttcg gcccgtaggc 150 gtctggcagc ccggcgccatcttcatcgag cgccatggcc gcagcctgcg 200 ggccgggagc ggccgggtac tgcttgctcctcggcttgca tttgtttctg 250 ctgaccgcgg gccctgccct gggctggaac gaccctgacagaatgttgct 300 gcgggatgta aaagctctta ccctccacta tgaccgctat accacctccc350 gcaggctgga tcccatccca cagttgaaat gtgttggagg cacagctggt 400tgtgattctt ataccccaaa agtcatacag tgtcagaaca aaggctggga 450 tgggtatgatgtacagtggg aatgtaagac ggacttagat attgcataca 500 aatttggaaa aactgtggtgagctgtgaag gctatgagtc ctctgaagac 550 cagtatgtac taagaggttc ttgtggcttggagtataatt tagattatac 600 agaacttggc ctgcagaaac tgaaggagtc tggaaagcagcacggctttg 650 cctctttctc tgattattat tataagtggt cctcggcgga ttcctgtaac700 atgagtggat tgattaccat cgtggtactc cttgggatcg cctttgtagt 750ctataagctg ttcctgagtg acgggcagta ttctcctcca ccgtactctg 800 agtatcctccattttcccac cgttaccaga gattcaccaa ctcagcagga 850 cctcctcccc caggctttaagtctgagttc acaggaccac agaatactgg 900 ccatggtgca acttctggtt ttggcagtgcttttacagga caacaaggat 950 atgaaaattc aggaccaggg ttctggacag gcttgggaactggtggaata 1000 ctaggatatt tgtttggcag caatagagcg gcaacaccct tctcagactc1050 gtggtactac ccgtcctatc ctccctccta ccctggcacg tggaataggg 1100cttactcacc ccttcatgga ggctcgggca gctattcggt atgttcaaac 1150 tcagacacgaaaaccagaac tgcatcagga tatggtggta ccaggagacg 1200 ataaagtaga aagttggagtcaaacactgg atgcagaaat tttggatttt 1250 tcatcacttt ctctttagaa aaaaagtactacctgttaac aattgggaaa 1300 aggggatatt caaaagttct gtggtgttat gtccagtgtagctttttgta 1350 ttctattatt tgaggctaaa agttgatgtg tgacaaaata cttatgtgtt1400 gtatgtcagt gtaacatgca gatgtatatt gcagtttttg aaagtgatca 1450ttactgtgga atgctaaaaa tacattaatt tctaaaacct gtgatgccct 1500 aagaagcattaagaatgaag gtgttgtact aatagaaact aagtacagaa 1550 aatttcagtt ttaggtggttgtagctgatg agttattacc tcatagagac 1600 tataatattc tatttggtat tatattatttgatgtttgct gttcttcaaa 1650 catttaaatc aagctttgga ctaattatgc taatttgtgagttctgatca 1700 cttttgagct ctgaagcttt gaatcattca gtggtggaga tggccttctg1750 gtaactgaat attaccttct gtaggaaaag gtggaaaata agcatctaga 1800aggttgttgt gaatgactct gtgctggcaa aaatgcttga aacctctata 1850 tttctttcgttcataagagg taaaggtcaa atttttcaac aaaagtcttt 1900 taataacaaa agcatgcagttctctgtgaa atctcaaata ttgttgtaat 1950 agtctgtttc aatcttaaaa agaatca 197780 339 PRT Homo Sapien 80 Met Ala Ala Ala Cys Gly Pro Gly Ala Ala GlyTyr Cys Leu Leu 1 5 10 15 Leu Gly Leu His Leu Phe Leu Leu Thr Ala GlyPro Ala Leu Gly 20 25 30 Trp Asn Asp Pro Asp Arg Met Leu Leu Arg Asp ValLys Ala Leu 35 40 45 Thr Leu His Tyr Asp Arg Tyr Thr Thr Ser Arg Arg LeuAsp Pro 50 55 60 Ile Pro Gln Leu Lys Cys Val Gly Gly Thr Ala Gly Cys AspSer 65 70 75 Tyr Thr Pro Lys Val Ile Gln Cys Gln Asn Lys Gly Trp Asp Gly80 85 90 Tyr Asp Val Gln Trp Glu Cys Lys Thr Asp Leu Asp Ile Ala Tyr 95100 105 Lys Phe Gly Lys Thr Val Val Ser Cys Glu Gly Tyr Glu Ser Ser 110115 120 Glu Asp Gln Tyr Val Leu Arg Gly Ser Cys Gly Leu Glu Tyr Asn 125130 135 Leu Asp Tyr Thr Glu Leu Gly Leu Gln Lys Leu Lys Glu Ser Gly 140145 150 Lys Gln His Gly Phe Ala Ser Phe Ser Asp Tyr Tyr Tyr Lys Trp 155160 165 Ser Ser Ala Asp Ser Cys Asn Met Ser Gly Leu Ile Thr Ile Val 170175 180 Val Leu Leu Gly Ile Ala Phe Val Val Tyr Lys Leu Phe Leu Ser 185190 195 Asp Gly Gln Tyr Ser Pro Pro Pro Tyr Ser Glu Tyr Pro Pro Phe 200205 210 Ser His Arg Tyr Gln Arg Phe Thr Asn Ser Ala Gly Pro Pro Pro 215220 225 Pro Gly Phe Lys Ser Glu Phe Thr Gly Pro Gln Asn Thr Gly His 230235 240 Gly Ala Thr Ser Gly Phe Gly Ser Ala Phe Thr Gly Gln Gln Gly 245250 255 Tyr Glu Asn Ser Gly Pro Gly Phe Trp Thr Gly Leu Gly Thr Gly 260265 270 Gly Ile Leu Gly Tyr Leu Phe Gly Ser Asn Arg Ala Ala Thr Pro 275280 285 Phe Ser Asp Ser Trp Tyr Tyr Pro Ser Tyr Pro Pro Ser Tyr Pro 290295 300 Gly Thr Trp Asn Arg Ala Tyr Ser Pro Leu His Gly Gly Ser Gly 305310 315 Ser Tyr Ser Val Cys Ser Asn Ser Asp Thr Lys Thr Arg Thr Ala 320325 330 Ser Gly Tyr Gly Gly Thr Arg Arg Arg 335

What is claimed is:
 1. Isolated nucleic acid having at least 80% nucleicacid sequence identity to a nucleotide sequence that encodes an aminoacid sequence selected from the group consisting of the amino acidsequence shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9), FIG. 6(SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18), FIG. 12(SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35), FIG. 18(SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50), FIG. 24(SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59), FIG. 30(SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65), FIG. 36(SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74), FIG. 42(SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) and FIG. 46 (SEQ ID NO:80). 2.Isolated nucleic acid having at least 80% nucleic acid sequence identityto a nucleotide sequence selected from the group consisting of thenucleotide sequence shown in FIG. 1 (SEQ ID NO:1), FIG. 3 (SEQ ID NO:8),FIG. 5 (SEQ ID NO:10), FIG. 7 (SEQ ID NO:15), FIG. 9 (SEQ ID NO:17),FIG. 11 (SEQ ID NO:22), FIG. 13 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:34),FIG. 17 (SEQ ID NO:39), FIG. 19 (SEQ ID NO:44), FIG. 21 (SEQ ID NO:49),FIG. 23 (SEQ ID NO:51), FIG. 25 (SEQ ID NO:56), FIG. 27 (SEQ ID NO:58),FIG. 29 (SEQ ID NO:60), FIG. 31 (SEQ ID NO:62), FIG. 33 (SEQ ID NO:64),FIG. 35 (SEQ ID NC:66), FIG. 37 (SEQ ID NO:71), FIG. 39 (SEQ ID NO:73),FIG. 41 (SEQ ID NO:75), FIG. 43 (SEQ ID NO:77) and FIG. 45 (SEQ IDNO:79).
 3. Isolated nucleic acid having at least 80% nucleic acidsequence identity to a nucleotide sequence selected from the groupconsisting of the full-length coding sequence of the nucleotide sequenceshown in FIG. 1 (SEQ ID NO:1), FIG. 3 (SEQ ID NO:8), FIG. 5 (SEQ IDNO:10), FIG. 7 (SEQ ID NO:15), FIG. 9 (SEQ ID NO:17), FIG. 11 (SEQ IDNO:22), FIG. 13 (SEQ ID NO:28), FIG. 15 (SEQ ID NO:34), FIG. 17 (SEQ IDNO:39), FIG. 19 (SEQ ID NO:44), FIG. 21 (SEQ ID NO:49), FIG. 23 (SEQ IDNO:51), FIG. 25 (SEQ ID NO:56), FIG. 27 (SEQ ID NO:58), FIG. 29 (SEQ IDNO:60), FIG. 31 (SEQ ID NO:62), FIG. 33 (SEQ ID NO:64), FIG. 35 (SEQ IDNO:66), FIG. 37 (SEQ ID NO:71), FIG. 39 (SEQ ID NO:73), FIG. 41 (SEQ IDNO:75), FIG. 43 (SEQ ID NO:77) and FIG. 45 (SEQ ID NO:79).
 4. Isolatednucleic acid having at least 80% nucleic acid sequence identity to thefull-length coding sequence of the DNA deposited under any ATCCaccession number shown in Table
 7. 5. A vector comprising the nucleicacid of any one of claims 1 to
 4. 6. The vector of claim 5 operablylinked to control sequences recognized by a host cell transformed withthe vector.
 7. A host cell comprising the vector of claim
 5. 8. The hostcell of claim 7, wherein said cell is a CHO cell.
 9. The host cell ofclaim 7, wherein said cell is an E. coli.
 10. The host cell of claim 7,wherein said cell is a yeast cell.
 11. A process for producing a PROpolypeptides comprising culturing the host cell of claim 7 underconditions suitable for expression of said PRO polypeptide andrecovering said PRO polypeptide from the cell culture.
 12. An isolatedpolypeptide having at least 80% amino acid sequence identity to an aminoacid sequence selected from the group consisting of the amino acidsequence shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9), FIG. 6(SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18), FIG. 12(SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35), FIG. 18(SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50), FIG. 24(SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59), FIG. 30(SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65), FIG. 36(SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74), FIG. 42(SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) and FIG. 46 (SEQ ID NO:80). 13.An isolated polypeptide scoring at least 80% positives when compared toan amino acid sequence selected from the group consisting of the aminoacid sequence shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9), FIG.6 (SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18), FIG. 12(SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35), FIG. 18(SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50), FIG. 24(SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59), FIG. 30(SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65), FIG. 36(SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74), FIG. 42(SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) and FIG. 46 (SEQ ID NO:80). 14.An isolated polypeptide having at least 80% amino acid sequence identityto an amino acid sequence encoded by the full-length coding sequence ofthe DNA deposited under any ATCC accession number shown in Table
 7. 15.A chimeric molecule comprising a polypeptide according to any one ofclaims 12 to 14 fused to a heterologous amino acid sequence.
 16. Thechimeric molecule of claim 15, wherein said heterologous amino acidsequence is an epitope tag sequence.
 17. The chimeric molecule of claim15, wherein said heterologous amino acid sequence is a Fc region of animmunoglobulin.
 18. An antibody which specifically binds to apolypeptide according to any one of claims 12 to
 14. 19. The antibody ofclaim 18, wherein said antibody is a monoclonal antibody, a humanizedantibody or a single-chain antibody.
 20. Isolated nucleic acid having atleast 80% nucleic acid sequence identity to: (a) a nucleotide sequenceencoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ IDNO:9), FIG. 6 (SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ IDNO:18), FIG. 12 (SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ IDNO:35), FIG. 18 (SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ IDNO:50), FIG. 24 (SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ IDNO:59), FIG. 30 (SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ IDNO:65), FIG. 36 (SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ IDNO:74), FIG. 42 (SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) or FIG. 46 (SEQID NO:80), lacking its associated signal peptide; (b) a nucleotidesequence encoding an extracellular domain of the polypeptide shown inFIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9), FIG. 6 (SEQ ID NO:11), FIG.8 (SEQ ID NO:16), FIG. 18 (SEQ ID NO:18), FIG. 12 (SEQ ID NO:23), FIG.14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35), FIG. 18 (SEQ ID NO:40), FIG.20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50), FIG. 24 (SEQ ID NO:52), FIG.26 (SEQ ID NC):57), FIG. 28 (SEQ ID NO:59), FIG. 30 (SEQ ID NO:61), FIG.32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65), FIG. 36 (SEQ ID NO:67), FIG.38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74), FIG. 42 (SEQ ID NO:76), FIG.44 (SEQ ID NO:78) or FIG. 46 (SEQ ID NO:80), with its associated signalpeptide; or (c) a nucleotide sequence encoding an extracellular domainof the polypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9),FIG. 6 (SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18),FIG. 12 (SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35),FIG. 18 (SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50),FIG. 24 (SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59),FIG. 30 (SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65),FIG. 36 (SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74),FIG. 42 (SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) or FIG. 46 (SEQ IDNO:80), lacking its associated signal peptide.
 21. An isolatedpolypeptide having at least 80% amino acid sequence identity to: (a) thepolypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9), FIG. 6(SEQ ID NO:1), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18), FIG. 12(SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35), FIG. 18(SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50), FIG. 24(SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59), FIG. 30(SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65), FIG. 36(SEQ ID NO:67), FIG. 38 (S.EQ ID NO:72), FIG. 40 (SEQ ID NO:74), FIG. 42(SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) or FIG. 46 (SEQ ID NO:80),lacking its associated signal peptide; (b) an extracellular domain ofthe polypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ ID NO:9),FIG. 6 (SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ ID NO:18),FIG. 12 (SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ ID NO:35),FIG. 18 (SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ ID NO:50),FIG. 24 (SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ ID NO:59),FIG. 30 (SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ ID NO:65),FIG. 36 (SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ ID NO:74),FIG. 42 (SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) or FIG. 46 (SEQ IDNO:80), with its associated signal peptide; or (c) an extracellulardomain of the polypeptide shown in FIG. 2 (SEQ ID NO:2), FIG. 4 (SEQ IDNO:9), FIG. 6 (SEQ ID NO:11), FIG. 8 (SEQ ID NO:16), FIG. 10 (SEQ IDNO:18), FIG. 12(SEQ ID NO:23), FIG. 14 (SEQ ID NO:29), FIG. 16 (SEQ IDNO:35), FIG. 18 (SEQ ID NO:40), FIG. 20 (SEQ ID NO:45), FIG. 22 (SEQ IDNO:50), FIG. 24 (SEQ ID NO:52), FIG. 26 (SEQ ID NO:57), FIG. 28 (SEQ IDNO:59), FIG. 30 (SEQ ID NO:61), FIG. 32 (SEQ ID NO:63), FIG. 34 (SEQ IDNO:65), FIG. 36 (SEQ ID NO:67), FIG. 38 (SEQ ID NO:72), FIG. 40 (SEQ IDNO:74), FIG. 42 (SEQ ID NO:76), FIG. 44 (SEQ ID NO:78) or FIG. 46 (SEQID NO:80), lacking its associated signal peptide.